Role of vit c in collagen byosynthesis

Collagen
Homeostasis
Bones- osteoblasts, osteoclasts
Elastin and keratin
Osteogenesis imperfect
SDS polyacrelamide gel

PBL1 Fragile bones: Handle with care

CASE HISTORY
A patient with a painful arm attends the casualty department
A teenage girl attended her local hospital casualty department complaining of severe pain in
her right arm. She explained to the House Officer that she had accidentally hit it, but rather
gently, on the side of her dressing table while picking up her pet cat. The pain was
excruciating. Her mother was contacted. Subsequent interview and examination revealed that
the girl had a history of multiple fractures and that these had fully healed in the past. Her foetal
development and birth appeared normal and she scored normally in the Apgar test. Her brother
and sister have normal medical histories.
Medical examination
The girl was normally developed for a mid-adolescent female but with a slightly short stature
for her age. Her rib cage was also slightly bowed. She had regular weight and physique for her
height. She was in obvious pain and had no use in her right arm. There was no evidence of
external body bruising or lacerations Head and neck examination showed that the normal outer
white coat of the eye (schlera) had a very pale bluish tinge. She had some hearing impairment
and her dentition was poor but mental functions were normal. Her heart and lung functions
were normal. The bones of her limbs were slightly bowed and her joints loose.
X-ray examination of the girl’s arms and legs revealed a transverse fracture of the right radius.
This examination also revealed some osteopenia. A follow-up X-ray of the head revealed
mottled areas consistent with loss of bone density probably caused by irregular ossification.
Clinical tests
A skin biopsy was taken with the patient’s consent.
Part of the sample was used for DNA sequencing.
An SDS-polyacrylamide gel electrophoresis of labelled skin fibroblast collagen is shown below
 C P C P

α1(I) α1(I)
α2(I) α2(I)

Reducing Non-reducing
conditions conditions

Key: C = control, normal patient, P = patient

Monolayer skin fibroblasts were grown from the remainder of the biopsy sample and the
proteins were labelled in the growing cells with [3H] proline and [35S] cysteine. The labelled
collagen secreted by the cells was extracted, solubilised and analysed by SDS-polyacrylamide
gel electrophoresis under both normal and mercaptoethanol treated conditions. The normal gel
from the patient (no mercaptoethanol), but not from the control gel, revealed an abnormal, low-
mobility band of type I collagen. This result is consistent with a point mutation in the COL1A1
gene. DNA sequence analysis confirmed the prediction.

Apgar Test
The very first test given to your newborn, the Apgar score occurs right after your baby's
birth in the delivery or birthing room. The test was designed to quickly evaluate a
newborn's physical condition after delivery and to determine any immediate need for extra
medical or emergency care. Although the Apgar score was developed in 1952 by an
anesthesiologist named Virginia Apgar, you may have also heard it referred to as an
acronym for: Activity, Pulse, Grimace,Appearance, and Respiration.

The Apgar test is usually given to your baby twice: once at 1 minute after birth, and again
at 5 minutes after birth. Rarely, if there are concerns about the baby's condition and the
first two scores are low, the test may be scored for a third time at 10 minutes after birth.

Five factors are used to evaluate the baby's condition and each factor is scored on a scale
of 0 to 2, with 2 being the best score:

1. activity and muscle tone

2. pulse (heart rate)
3. grimace response (medically known as "reflex irritability")
4. appearance (skin coloration)
5. respiration (breathing rate and effort)
Doctors, midwives, or nurses add these five factors together to calculate the Apgar score.
Scores obtainable are between 10 and 0, with 10 being the highest possible score.

Apgar Scoring
Apgar Sign 2 1 0
Heart Rate Normal (above Below 100 Absent
(pulse) 100 beats per beats per (no pulse)
minute) minute
Breathing Normal rate Slow or Absent (no
(rate and effort) and effort, irregular breathing)
good cry breathing,
weak cry
Grimace(responsiveness Pulls away, Facial Absent (no
or "reflex irritability") sneezes, or movement response to
coughs with only stimulation)
stimulation (grimace)
with
stimulation
Activity Active, Arms and No
(muscle tone) spontaneous legs flexed movement,
movement with little "floppy" tone
movement
Appearance Normal color Normal color Bluish-gray or
(skin coloration) all over (but hands pale all over
(hands and and feet are
feet are pink) bluish)

A baby who scores a 7 or above on the test at 1 minute after birth is generally considered
in good health. However, a lower score doesn't necessarily mean that your baby is
unhealthy or abnormal. But it may mean that your baby simply needs some special
immediate care, such as suctioning of the airways or oxygen to help him or her breathe,
after which your baby may improve.

At 5 minutes after birth, the Apgar score is recalculated, and if your baby's score hasn't
improved to 7 or greater, or there are other concerns, the doctors and nurses may
continue any necessary medical care and will closely monitor your baby. Some babies are
born with heart or lung conditions or other problems that require extra medical care;
others just take a little longer than usual to adjust to life outside the womb. Most
newborns with initial Apgar scores of less than 7 will eventually do just fine.

It's important for new parents to keep their baby's Apgar score in perspective. The test
was designed to help health care providers assess a newborn's overall physical condition
so that they could quickly determine whether the baby needed immediate medical care. It
was not designed to predict a baby's long-term health, behavior, intellectual status, or
outcome. Few babies score a perfect 10, and perfectly healthy babies sometimes have a
lower-than-usual score, especially in the first few minutes after birth.
Keep in mind that a slightly low Apgar score (especially at 1 minute) is normal for some
newborns, especially those born after a high-risk pregnancy, cesarean section, or a
complicated labor and delivery. Lower Apgar scores are also seen in premature babies,
who usually have less muscle tone than full-term newborns and who, in many cases, will
require extra monitoring and breathing assistance because of their immature lungs.

If your doctor or midwife is concerned about your baby's score, he or she will let you know
and will explain how your baby is doing, what might be causing problems, if any, and what
care is being given. For the most part, though, most babies do very well, so relax and
enjoy the moment!

The term laceration implies a torn or jagged wound. Lacerations tend to be caused by
blunt trauma (such as a blow, fall, or collision). Cuts and lacerations are terms for the same
condition.

Osteopenia is a condition where bone mineral density is lower than normal. It is considered
by many doctors to be a precursor to osteoporosis. However, not every person diagnosed
with osteopenia will develop osteoporosis. More specifically, osteopenia is defined as a bone
mineral density T-score between -1.0 and -2.5.

Osteopenia was defined in June 1992 by the World Health Organization. A group of experts
decided that condition would mean a bone density that was one standard deviation below that
of an average 30-year-old white woman. The group also defined osteoporosis as bone
density 2.5 standard deviations or more below that 30-year-old;[2] previously it had been used
only in cases where elderly patients had fractured or broken a bone.[3] An osteoporosis
epidemiologist at the Mayo Clinic who participated in setting the criteria in 1992 said "It was
just meant to indicate the emergence of a problem," and noted that "It didn't have any
particular diagnostic or therapeutic significance. It was just meant to show a huge group who
looked like they might be at risk."[2]

The definition has been controversial. Steven R. Cummings, of the University of California,
San Francisco, said in 2003 that "There is no basis, no biological, social, economic or
treatment basis, no basis whatsoever" for using one standard deviation. Cummings added
that "As a consequence, though, more than half of the population is told arbitrarily that they
have a condition they need to worry about."
Osteopenia: Mild thinning of the bone mass, but not as severe as osteoporosis. Osteopenia
results when the formation of bone (osteoid synthesis) is not enough to offset normal bone
loss (bone lysis). Osteopenia is generally considered the first step along the road to
osteoporosis, a serious condition in which bone density is extremely low and bones are
porous and prone to shatter. Diminished bone calcification, as seen on plain X-ray film, is
referred to as osteopenia, whether or not osteoporosis is present. The diagnosis of
osteopenia may also be made by a special X-ray machine for bone density testing.

The World Health Organization (WHO) recognizes osteopenia, for people 50 and older with
lower than average bone density who do not have osteoporosis. WHO defines osteopenia as
a bone density between one standard deviation (SD) and 2.5 SD below the bone density of a
normal young adult. (Osteoporosis is defined as 2.5 SD or more below that reference point.)

"Osteopenia" has two Greek ancestors: "osteon", bone and "penia", poverty = bone poverty.

Ossification (or osteogenesis) is the process of laying down new bone material by cells
called osteoblasts. It is synonymous with bone tissue formation. There are two processes
resulting in the formation of normal, healthy bone tissue:[1] Intramembranous ossification is the
direct laying down of bone into the primitive connective tissue (mesenchyme),
while endochondral ossification involves cartilage as a precursor. In fracture healing,
endochondral osteogenesis is the most commonly occurring process, for example in fractures
of long bones treated by plaster of Paris, whereas fractures treated by open reduction and
stabilization by metal plate and screws may heal by intramembranous osteogenesis.

Heterotopic ossification is a process resulting in the formation of bone tissue that is often
atypical, at an extraskeletal location. Calcification is often confused with ossification.
Calcification is synonymous with the formation of calcium-based salts and crystals within cells
and tissue. It is a process that occurs during ossification, but not vice versa.

The exact mechanisms by which bone development is triggered remains unclear, but it
involves growth factors and cytokines in some way.

Skin biopsy is a biopsy technique in which a skin lesion is removed and sent to
the pathologist to render a microscopic diagnosis. It is usually done under local anesthetic in
a physician's office, and results are often available in 4 to 10 days. It is commonly performed
by dermatologists.
A skin biopsy is a procedure in which a sample of skin tissue is removed, processed, and
examined under a microscope.

Several different methods may be used to obtain a skin sample, depending on the size and
location of the abnormal area of skin, called a skin lesion. The skin sample is placed in a
solution, such as formaldehyde, or in a sterile container if infection is suspected. In each of
these procedures, the tissue is processed and then examined under a microscope.

Skin biopsies most often are done to diagnose skin cancer, which may be suspected when an
abnormal area of skin has changed color , shape , size, or appearance or has not
healed after an injury. Skin cancers are the most common type of cancers.
Early diagnosis of a suspicious skin lesion and skin biopsy can help identify skin cancers and
lead to early treatment.

http://en.wikipedia.org/wiki/SDS-PAGE

The purpose of SDS-PAGE is to separate proteins according to their size, and no other physical feature.
In order to understand how this works, we have to understand the two halves of the
name: SDS and PAGE.

SDS
Since we are trying to separate many different protein molecules of different shapes and sizes, we first
want to denatured so that the proteins no longer have any secondary, tertiary or quaternary structure
(i.e. we want them to retain only their primary amino acid structure). Consider two proteins that are
each 500 amino acids long but one is shaped like a closed umbrella whle the other one looks like an
open umbrella. If you tried to run down the street with both of these molecules under your arms, which
one would be more likely to slow you down, even though they weigh exactly the same? This analogy
illustrates mass and the 3D structure of a molecule will detrmine how well it can move through an
environment. We use SDS to denature all proteins to the same linear shape.

Figure 1. This cartoon depicts what happens to a protein (pink line) when it is incubated with the
denaturing detergent SDS. The top portion of the figure shows a protein with negative and positive
charges due to the charged R-groups in the protein. The large H's represent hydrophobic domains
where nonpolar R-groups have collected in an attempt to get away from the polar water that surrounds
the protein. The lower diagram shows that SDS can disrupt hydrophobic areas and coat proteins with
many negative charges which overwhelms any positive charges the protein had due to positively
charged R-groups. The resulting protein has been denatured by SDS (reduced to its primary structure)
and as a result has been linearized.

SDS (sodium dodecyl sulfate) is a detergent (soap) that can dissolve hydrophobic molecules but also
has a negative charge (sulfATE) attached to it. Therefore, if a cell is incubated with SDS, the
membranes will be dissolved, all the proteins will be soluablized by the detergent, plus all the proteins
will be covered with many negative charges. So a protein that started out like the one shown in the top
part of figure 1 will be converted into the one shown in the bottom part of figure 1. The end result has
two important features: 1) all proteins retain only their primary structure and 2) all proteins have a large
negative charge which means they will all migrate towards the positve pole when placed in an electric
field. Now we are ready to focus on the second half - PAGE.
PAGE
If the proteins are denatured and put into an electric field, they will all move towards the positive pole
at the same rate, with no separation by size. So we need to put the proteins into an environment that
will allow different sized proteins to move at different rates. The environment of choice is
polyacrylamide, which is a polymer of acrylamide monomers. When this polymer is formed, it turns
into a gel and we will use electricity to pull the proteins through the gel so the entire process is
called polyacrylamide gel electrophoresis (PAGE). A polyacrylamide gel is not solid but is made of a
laberynth of tunnels through a meshwork of fibers (figure 2 and figure 3).

Figure 2. This cartoon shows a slab of polyacrylamide (dark gray) with tunnels (different sized red
rings with shading to depict depth) exposed on the edge. Notice that there are many different sizes of
tunnels scattered randomly throughout the gel.

Figure 3. This is a top view of two selected tunnels (only two are shown for clarity of the diagram).
These tunnels extend all the way through the gel, but they meander through the gel and do not go in
straight lines. Notice the difference in diameter of the two tunnels.

Now we are ready to apply the mixture of denatured proteins to the gel and apply the current (figure 4).
If all the proteins enter the gel at the same time and have the same force pulling them towards the other
end, which ones will be able to move through the gel faster? Think of the gel as a tiny forrest with
many branches and twigs througout the forest but they form tunnels of different sizes. If we let children
and adults run through this forest at the same time, who will be able to get through faster? The children
of course. Why? Because of their small size, they move through the forest faster since they have access
to more of the paths in the forest while adults are limited to only the larger paths. Likewise, small
molecules can manuver through the polyacrylamide forest faster than big molecules.

Figure 4. Cartoon showing a mixutre of denatured proteins (pink lines of differen lengths) beginning
their journey through a polyacrylamide gel (gray slab with tunnels). An electric filed is established with
the positive pole (red plus) at the far end and the negative pole (black minus) at the closer end. Since all
the proteins have strong negative charges, they will all move in the direction the arrow is pointing (run
to red).

You have to remember that when we work with proteins, we work with many copies of each type of
protein. As a result, the collection of proteins of any given size tend to move through the gel at the
same rate, even if they do not take exactly the same tunnels to get through. Back to our analogy of the
forest... If we were in a hot air ballon above the forest and watched 100 children, 100 teenagers, and
100 large adults running through the forest, we would see a collection (or band) of children moving
quickly though each individual would choose his or her own route through the forrest. Behind the
smallest individuals, we would see a band of teenagers moving slower, and a third band made of adults
plodding their way through the forest using only the largest of paths. Likewise, proteins tend to move
through a gel in bunches, or bands, since there are so many copies of each protein and they are all the
same shape and size. When running an SDS-PAGE, we never let the proteins electrophorese (run) so
long that they actually reach the other side of the gel. We turn off the current and then stain the
proteins and see how far they moved through the gel (until we stain them, they are colorless and thus
invisible). Figure 5 shows a cartoon gel and figre 6 shows a real one. Notice that the actual bands are
equal in size, but the proteins within each band are of different sizes.
Figure 5. Top view of an SDS PAGE after the current has been on for a while (positive pole at the
bottom) and then turned off. The gel (gray box) has five numbered lanes where five different samples
of proteins (many copies of each kind of protein) were applied to the gel. (Lane 1, molecular weight
standards of known sizes; Lane 2, a mixture of three proteins of different sizes with a being the largest
and c being the smallest protein; Lane 3, protein a by itself; Lane 4, protein b by itself; Lane 5
protein c by itself.) Notice that each group of the three proteins migrated the same distance in the gel
whether they were with other proteins (lane 2) or not (lanes 3-5). The molecular weight standards are
used to measure the relative sizes of the unknow proteins (a, b, and c).

Figure 6. This photo shows a variety of different proteins being separated on a gel. This particular
image is showing a serial dilution of the same protein sample to indicate how little protein is needed
(16 picograms = 16 . 10 -12 grams) in order to be detected.
This image was taken from a home page operated by Hitachi Software.
There is a caveot to this method that you must always keep in mind. SDS-PAGE separates proteins
based on their primary structure or size but not amino acid sequence. Therefore, if we had many copies
of two different proteins that were both 500 amino acids long, they would travel together through the
gel in a mixed band. As a result, we would not be able to use SDS-PAGE to separate these two proteins
of the same molecular weight from each other.

A fibroblast is a type of cell that synthesizes the extracellular matrix and collagen,[1] the
structural framework (stroma) for animal tissues, and plays a critical role in wound healing.
Fibroblasts are the most common cells of connective tissue in animals. Fibroblasts
and fibrocytes are two states of the same cells, the former being the activated state, the latter
the less active state, concerned with maintenance and tissue metabolism. Currently, there is
a tendency to call both forms fibroblasts. The suffix "blast" is used in cellular biology to denote
a stem cell or a cell in an activated state of metabolism.

Fibroblasts are morphologically heterogeneous with diverse appearances depending on their

location and activity. Though morphologically inconspicuous, ectopically transplanted
fibroblasts can often retain positional memory of the location and tissue context where they
had previously resided, at least over a few generations. This remarkable behavior may lead to
discomfort in the rare event that they stagnate there excessively.

Embryologic origin
The main function of fibroblasts is to maintain the structural integrity of connective tissues by
continuously secreting precursors of the extracellular matrix. Fibroblasts secrete the
precursors of all the components of the extracellular matrix, primarily the ground
substance and a variety of fibers. The composition of the extracellular matrix determines the
physical properties of connective tissues.

Like other cells of connective tissue, fibroblasts are derived from primitive mesenchyme. Thus
they express the intermediate filament protein vimentin, a feature used as a marker to
distinguish theirmesodermal origin. However, this test is not specific as epithelial cells
cultured in vitro on adherent substratum may also express vimentin after some time.

In certain situations epithelial cells can give rise to fibroblasts, a process called epithelial-
mesenchymal transition (EMT).

Conversely, fibroblasts in some situations may give rise to epithelia by undergoing

a mesenchymal to epithelial transition (MET) and organizing into a condensed, polarized,
laterally connected true epithelial sheet. This process is seen in many developmental
situations (e.g. nephron and notocord development).
Fibroblasts have a branched cytoplasm surrounding an elliptical, speckled nucleus having
one or two nucleoli. Active fibroblasts can be recognized by their abundant
rough endoplasmic reticulum. Inactive fibroblasts, which are also called fibrocytes, are
smaller and spindle shaped. They have a reduced rough endoplasmic reticulum.
Althoughdisjointed and scattered when they have to cover a large space, fibroblasts when
crowded often locally align in parallel clusters.

Fibroblasts make collagens, glycosaminoglycans, reticular and elastic

fibers, glycoproteins found in the extracellular matrix and cytokine TSLP. Growing individuals'
fibroblasts are dividing and synthesizing ground substance. Tissue damage stimulates
fibrocytes and induces the mitosis of fibroblasts.

Unlike the epithelial cells lining the body structures, fibroblasts do not form flat monolayers
and are not restricted by a polarizing attachment to a basal lamina on one side, although they
may contribute to basal lamina components in some situations (e.g.
subepithelial myofibroblasts in intestine may secrete the α-2 chain carrying component of
the laminin which is absent only in regions of follicle associated epithelia which lack the
myofibroblast lining). Fibroblasts can also migrate slowly over substratum as individual cells,
again in contrast to epithelial cells. While epithelial cells form the lining of body structures, it is
fibroblasts and related connective tissues which sculpt the "bulk" of an organism.

The life span of a fibroblast, as measured in chick embryos, is 57 ± 3 days.[2

Collagen (English pronunciation: /ˈkɒlədʒən/) is a group of naturally occurring proteins. In nature,

it is found exclusively in animals, especially in the flesh and connective tissues ofmammals.
[1]
It is the main component of connective tissue, and is the most abundant protein in
mammals,[2] making up about 25% to 35% of the whole-body protein content. Collagen, in the
form of elongated fibrils, is mostly found in fibrous tissues such as tendon, ligament and skin,
and is also abundant in cornea, cartilage, bone, blood vessels, the gut, and intervertebral
disc.

In muscle tissue it serves as a major component of endomysium. Collagen constitutes 1% to

2% of muscle tissue, and accounts for 6% of the weight of strong, tendinous muscles.
[3]
Gelatin, which is used in food and industry, is collagen that has been
irreversibly hydrolyzed.

History and background

The molecular and packing structures of collagen have eluded scientists over decades of
research. The first evidence that it possesses a regular structure at the molecular level was
presented in the mid-1930s.[4][5] Since that time many prominent scholars, including Nobel
laureates Crick, Pauling, Rich and Yonath and others including Brodsky, Berman,
and Ramachandran, concentrated on the conformation of the collagen monomer. Several
competing models, although correctly dealing with the conformation of each individual peptide
chain, gave way to the triple-helical "Madras" model which provided an essentially correct
model of the molecule's quaternary structure[6][7][8] although this model still required some
refinement.[9][10][11][12] The packing structure of collagen has not been defined to the same
degree outside of the fibrillar collagen types, although it has been long known to be
hexagonal ...or quasi-hexagonal.[13][14][15] As with its monomeric structure, several conflicting
models alleged that either the packing arrangement of collagen molecules is 'sheet-like'
or microfibrillar.[16][17] The microfibrillar structure of collagen fibrils in tendon, cornea and
cartilage has been directly imaged by electron microscopy.[18][19][20] In 2006, it was confirmed
that the microfibrillar structure of adult tendon as described by Fraser, Miller, Wess (amongst
others) was closest to the observed structure, although it over-simplified the topological
progression of neighboring collagen molecules and hence did not predict the correct
conformation of the discontinuous D-periodic pentameric arrangement termed simply: the
microfibril.[21]

[edit]Molecular structure
The tropocollagen or "collagen molecule" is a subunit of larger collagen aggregates such as
fibrils. It is approximately 300 nm long and 1.5 nm in diameter, made up of
three polypeptide strands (called alpha chains), each possessing the conformation of a left-
handed helix (its name is not to be confused with the commonly occurring alpha helix, a right-
handed structure). These three left-handed helices are twisted together into a right-
handed coiled coil, a triple helix or "super helix", a cooperative quaternary structure stabilized
by numerous hydrogen bonds. With type I collagen and possibly all fibrillar collagens if not all
collagens, each triple-helix associates into a right-handed super-super-coil that is referred to
as the collagen microfibril. Each microfibril is interdigitated with its neighboring microfibrils to a
degree that might suggest that they are individually unstable although within collagen fibrils
they are so well ordered as to be crystalline.

A distinctive feature of collagen is the regular arrangement of amino acids in each of the three
chains of these collagen subunits. The sequence often follows the pattern Gly-Pro-X or Gly-X-
Hyp, where X may be any of various other amino acid residues. Proline or hydroxyproline
constitute about 1/6 of the total sequence. With glycine accounting for the 1/3 of the
sequence, this means that approximately half of the collagen sequence is not glycine, proline
or hydroxyproline, a fact often missed due to the distraction of the unusual GX1X2 character of
collagen alpha-peptides. This kind of regular repetition and high glycine content is found in
only a few other fibrous proteins, such as silk fibroin. About 75-80% of silk is (approximately)
-Gly-Ala-Gly-Ala- with 10% serine, and elastin is rich in glycine, proline, and alanine (Ala),
whose side group is a small, inert methyl group. Such high glycine and regular repetitions are
never found in globular proteins save for very short sections of their sequence. Chemically-
reactive side groups are not needed in structural proteins as they are in enzymes
and transport proteins, however collagen is not quite just a structural protein. Due to its key
role in the determination of cell phenotype, cell adhesion, tissue regulation and infrastructure,
many sections of its non-proline rich regions have cell or matrix association / regulation roles.
The relatively high content of proline and hydroxyproline rings, with their geometrically
constrained carboxyl and (secondary) amino groups, along with the rich abundance of
glycine, accounts for the tendency of the individual polypeptide strands to form left-handed
helices spontaneously, without any intrachain hydrogen bonding.

Because glycine is the smallest amino acid with no side chain, it plays a unique role in fibrous
structural proteins. In collagen, Gly is required at every third position because the assembly of
the triple helix puts this residue at the interior (axis) of the helix, where there is no space for a
larger side group than glycine’s single hydrogen atom. For the same reason, the rings of the
Pro and Hyp must point outward. These two amino acids help stabilize the triple helix—Hyp
even more so than Pro; a lower concentration of them is required in animals such as fish,
whose body temperatures are lower than most warm-blooded animals.

[edit]Fibrillar structure
The tropocollagen subunits spontaneously self-assemble, with regularly staggered ends, into
even larger arrays in the extracellular spaces of tissues.[22][23] In the fibrillar collagens, the
molecules are staggered from each other by about 67 nm (a unit that is referred to as ‘D’ and
changes depending upon the hydration state of the aggregate). Each D-period contains 4 and
a fraction collagen molecules. This is because 300 nm divided by 67 nm does not give an
integer (the length of the collagen molecule divided by the stagger distance D). Therefore in
each D-period repeat of the microfibril, there is a part containing five molecules in cross-
section—called the “overlap” and a part containing only 4 molecules, called the "gap".[21] The
triple-helices are also arranged in a hexagonal or quasi-hexagonal array in cross-section, in
both the gap and overlap regions.[13][21]

There is some covalent crosslinking within the triple helices, and a variable amount of
covalent crosslinking between tropocollagen helices forming well organized aggregates (such
as fibrils).[24] Larger fibrillar bundles are formed with the aid of several different classes of
proteins (including different collagen types), glycoproteins and proteoglycans to form the
different types of mature tissues from alternate combinations of the same key players.
[23]
Collagen's insolubility was a barrier to the study of monomeric collagen until it was found
that tropocollagen from young animals can be extracted because it is not yet fully crosslinked.
However, advances in microscopy techniques electron microscopy (EM) and atomic force
microscopy (AFM)) and X-ray diffraction have enabled researchers to obtain increasingly
 detailed images of collagen structure in situ. These later advances are particularly important
to better understanding the way in which collagen structure affects cell-cell and cell-matrix
communication, and how tissues are constructed in growth and repair, and changed in
development and disease.[25][26] For example using AFM –based nanoindentation it has been
shown that a single collagen fibril is a heterogeneous material along its axial direction with
significantly different mechanical properties in its gap and overlap regions, correlating with its
different molecular organizations in these two regions.[27]

Collagen fibrils are semicrystalline aggregates of collagen molecules. Collagen fibers are
bundles of fibrils.
Collagen fibrils/aggregates are arranged in different combinations and concentrations
in various tissues to provide varying tissue properties. In bone, entire collagen triple
helices lie in a parallel, staggered array. Forty nm gaps between the ends of the
tropocollagen subunits (approximately equal to the gap region) probably serve as
nucleation sites for the deposition of long, hard, fine crystals of the mineral
component, which is (approximately) hydroxyapatite, Ca10(PO4)6(OH)2 with
some phosphate. It is in this way that certain kinds of cartilage turn into bone. Type I
collagen gives bone its tensile strength.
[edit]Types and associated disorders
Collagen occurs in many places throughout the body. So far, only 29 types of collagen have
been identified and described. Over 90% of the collagen in the body, however, is of type I[28]

 Collagen One: skin, tendon, vascular, ligature, organs, bone (main component of
bone)

 Collagen Two: cartilage (main component of cartilage)

 Collagen Three: reticulate (main component of reticular fibers), commonly found

alongside type I.

 Collagen Four: forms bases of cell basement membrane

 Collagen Five: cells surfaces, hair and placenta

Collagen-related diseases most commonly arise from genetic defects or nutritional

deficiencies that affect the biosynthesis, assembly, postranslational modification, secretion, or
other processes involved in normal collagen production.

Type Notes Gene(s) Disorders

I This is the most abundant COL1A1, COL1A2 osteogenesis

collagen of the human body. It imperfecta, Ehlers-
is present in scar tissue, the end Danlos
product when tissueheals by Syndrome, Infantile
repair. It is found in tendons, cortical
 skin, artery walls,
the endomysium of myofibrils, hyperostosis aka
fibrocartilage, and the organic Caffey's disease
part of bones and teeth.

Hyaline cartilage, makes up

50% of all cartilage Collagenopathy,
II COL2A1
protein. Vitreous humour of types II and XI
the eye.

This is the collagen

of granulation tissue, and is
produced quickly by young Ehlers-Danlos
fibroblasts before the tougher
III COL3A1 Syndrome, Dupuytr
type I collagen is
en's contracture
synthesized. Reticular fiber.
Also found in artery walls,
skin, intestines and the uterus

basal lamina; eye lens. Also

serves as part of the filtration Alport
COL4A1, COL4A2,COL4A3, COL4A4,COL4
IV system in capillaries and syndrome, Goodpast
A5, COL4A6
the glomeruli of nephron in ure's syndrome
thekidney.

most interstitial tissue, assoc. Ehlers-Danlos

V with type I, associated COL5A1, COL5A2,COL5A3 syndrome (Classical
with placenta )

Ulrich
most interstitial tissue, assoc.
VI COL6A1, COL6A2,COL6A3 myopathy and Bethl
with type I
em myopathy

forms anchoring
epidermolysis
VII fibrils in dermal epidermal COL7A1
bullosa dystrophica
junctions

Posterior
VIII some endothelial cells COL8A1, COL8A2 polymorphous
corneal dystrophy 2
 FACIT collagen, cartilage,
IX assoc. with type II and XI COL9A1, COL9A2,COL9A3 - EDM2 and EDM3
fibrils

hypertrophic and mineralizing COL10A1 Schmid metaphyseal

X
cartilage dysplasia

Collagenopathy,
XI cartilage COL11A1, COL11A2 types II and XI

FACIT collagen, interacts with

type I containing
XII COL12A1 -
fibrils, decorin and
glycosaminoglycans

transmembrane collagen,
interacts with integrin
a1b1, fibronectin and
XIII COL13A1 -
components of basement
membranes
likenidogen and perlecan.

XIV FACIT collagen COL14A1 -

XV - COL15A1 -

XVI - COL16A1 -

Bullous
transmembrane collagen, also pemphigoid and
XVII known as BP180, a 180 kDa COL17A1 certain forms of
protein junctional epidermol
ysis bullosa

XVIII source of endostatin COL18A1 -

XIX FACIT collagen COL19A1 -

XX - COL20A1 -
XXI FACIT collagen COL21A1 -

XXII - COL22A1 -

XXIII MACIT collagen - COL23A1 -

XXI
- COL24A1 -
V

XXV - COL25A1 -

XXV
- EMID2 -
I

XXV
- COL27A1 -
II

XXV
- COL28A1 -
III

XXI
epidermal collagen COL29A1 Atopic dermatitis[29]
X

In addition to the above mentioned disorders, excessive deposition of collagen occurs

in scleroderma.

[edit]Staining

In histology, collagen is brightly eosinophilic (pink) in standard H&E slides. The dye methyl
violet may be used to stain the collagen in tissue samples.

The dye methyl blue can also be used to stain collagen and immunohistochemical stains are
available if required.

The best stain for use in differentiating collagen from other fibers is Masson's trichrome stain.
[edit]Synthesis

Action of lysyl oxidase

[edit]Amino acids

Collagen has an unusual amino acid composition and sequence:

 Glycine (Gly) is found at almost every third residue

 Proline (Pro) makes up about 17% of collagen

 Collagen contains two uncommon derivative amino acids not directly inserted
during translation. These amino acids are found at specific locations relative to glycine
and are modified post-translationally by different enzymes, both of which require vitamin
C as a cofactor.

 Hydroxyproline (Hyp), derived from proline.

 Hydroxylysine (Hyl), derived from lysine (Lys). Depending on the type of

collagen, varying numbers of hydroxylysines are glycosylated (mostly
havingdisaccharides attached).

Cortisol stimulates degradation of (skin) collagen into amino acids.[30]

[edit]Collagen I formation

Most collagen forms in a similar manner, but the following process is typical for type I:
 1. Inside the cell

1. Two types of peptide chains are formed during translation on

ribosomes along the rough endoplasmic reticulum (RER): alpha-1 and
alpha-2 chains. These peptide chains (known as preprocollagen)
have registration peptides on each end and a signal peptide.

2. Polypeptide chains are released into the lumen of the RER.

3. Signal peptides are cleaved inside the RER and the chains are now
known as pro-alpha chains.

4. Hydroxylation of lysine and proline amino acids occurs inside the

lumen. This process is dependent on ascorbic acid (Vitamin C) as
a cofactor.

5. Glycosylation of specific hydroxylysine residues occurs.

6. Triple helical structure is formed inside the endoplasmic reticulum

from each two alpha-1 chains and one alpha-2 chain.

7. Procollagen is shipped to the golgi apparatus, where it is packaged

and secreted by exocytosis.

2. Outside the cell

1. Registration peptides are cleaved and tropocollagen is formed

by procollagen peptidase.

2. Multiple tropocollagen molecules form collagen fibrils, via covalent

cross-linking (aldol reaction) by lysyl oxidase which links hydroxylysine and
lysine residues. Multiple collagen fibrils form into collagen fibers.

3. Collagen may be attached to cell membranes via several types of

protein, including fibronectin and integrin.
[edit]Synthetic pathogenesis

Vitamin C deficiency causes scurvy, a serious and painful disease in which defective collagen
prevents the formation of strong connective tissue. Gums deteriorate and bleed, with loss of
teeth; skin discolors, and wounds do not heal. Prior to the eighteenth century, this condition
was notorious among long duration military, particularly naval, expeditions during which
participants were deprived of foods containing Vitamin C.

An autoimmune disease such as lupus erythematosus or rheumatoid arthritis[31] may attack

healthy collagen fibers.
Many bacteria and viruses have virulence factors which destroy collagen or interfere with its
production.

[edit]Use

Collagen is one of the long, fibrous structural proteins whose functions are quite different from
those of globular proteins such as enzymes. Tough bundles of collagen called collagen
fibers are a major component of the extracellular matrix that supports most tissues and gives
cells structure from the outside, but collagen is also found inside certain cells. Collagen has
great tensile strength, and is the main component
of fascia, cartilage, ligaments, tendons, bone and skin.[32][33] Along with soft keratin, it is
responsible for skin strength and elasticity, and its degradation leads to wrinkles that
accompany ageing[34][35]. It strengthens blood vessels and plays a role in tissue development.
It is present in the cornea and lens of the eye in crystalline form. It is also used in cosmetic
surgery andburns surgery. Hydrolyzed collagen can play an important role in weight
management, as a protein, it can be advantageously used for its satiating power.

Elastin is a protein in connective tissue that is elastic and allows many tissues in the body to
resume their shape after stretching or contracting. Elastin helps skin to return to its original
position when it is poked or pinched. Elastin is also an important load-bearing tissue in the
bodies of mammals and used in places where mechanical energy is required to be stored. In
humans, elastin is encoded by the ELN gene
Function
This gene encodes a protein that is one of the two components of elastic fibers. The encoded
protein is rich in hydrophobic amino acids such asglycine and proline, which form mobile
hydrophobic regions bounded by crosslinks between lysine residues.[2]

[edit]Clinical significance
Deletions and mutations in this gene are associated with supravalvular aortic stenosis (SVAS)
and autosomal dominant cutis laxa.[2]
Other associated defects in elastin include Marfan's Syndrome and emphysema caused by
alpha-1-antitrypsin deficiency.

[edit]Composition

Elastic fiber is composed of the protein fibrillin and elastin made of simple amino acids such
as glycine, valine, alanine, and proline.[3]

Elastin is made by linking many soluble tropoelastin protein molecules, in a

reaction catalyzed by lysyl oxidase, to make a massive insoluble, durable cross-linked array.
The amino acid responsible for these cross-links is lysine. Tropoelastin is a specialized
protein with a molecular weight of 64 to 66 kDa, and an irregular or random coil conformation
made up of 830 amino acids.
Desmosine and isodesmosine are types of links for the tropoelastin molecules.

[edit]Tissue distribution
Elastin serves an important function in arteries as a medium for pressure wave propagation to
help blood flow and is particularly abundant in large elastic blood vessels such as the aorta.
Elastin is also very important in the lungs, elastic ligaments, the skin, and the bladder, elastic
cartilage. It is present in all vertebrates above the jawless fish

Elastin is a protein found in the skin and tissue of the body. It helps to keep skin flexible but
tight, providing a bounce-back reaction if skin is pulled. Enough elastin in the skin means that
the skin will return to its normal shape after a pull. It also helps keep skin smooth as it
stretches to accommodate normal activities like flexing a muscle or opening and closing the
mouth to talk or eat.

Elastin tends to deplete as people age, resulting in wrinkled or stretched out skin. One might
note the “pregnancy pouch” many women have many years after having a baby. In part, the
leftover skin is a result of inadequate elastin, and also overstretching of the skin covering
theabdomen during pregnanc

Keratin refers to a family of fibrous structural proteins. Keratin is the key structural material
making up the outer layer of human skin. It is also the key structural component
of hair and nails. Keratin monomers assemble into bundles to form intermediate filaments,
which are tough and insoluble and form strong unmineralized tissues found
in reptiles, birds, amphibians, and mammals. The only other biological matter known to
approximate the toughness of keratinized tissue is chitin

Function
Keratin filaments are abundant in keratinocytes in the cornified layer of the epidermis; these
are cells which have undergone keratinization.

 the α-keratins in the hair (including wool), horns, nails, claws and hooves of
mammals[verification needed]

 the harder β-keratins found in nails and in the scales and claws of reptiles,
their shells (chelonians, such as tortoise, turtle, terrapin), and in the feathers, beaks,
claws of birds and quills of porcupines.[1] (These keratins are formed primarily in beta
sheets. However, beta sheets are also found in α-keratins.)[2]

The baleen plates of filter-feeding whales are made of keratin.

Although it is now difficult to be certain, the scales, claws, some protective armour and the
beaks of dinosaurs were probably composed of keratin.[3]

Keratins (also described as cytokeratins) are polymers of type I and type II intermediate
filaments, which have only been found in the genomes of chordates (vertebrates, Amphioxus,
urochordates). Nematodes and many other non-chordate animals seem to only have type
V intermediate filaments, lamins, which have a long rod domain (vs. a short rod domain for
the keratins).

[edit]Molecular biology and biochemistry

The usefulness of keratins depends on their supermolecular aggregation. These depend on
the properties of the individual polypeptide strands, which depend in turn on their amino
acid composition and sequence. The α-helix and β-sheet motifs, and disulfide bridges, are
crucial to the conformations of globular, functional proteins like enzymes, many of which
operate semi-independently, but they take on a completely dominant role in the architecture
and aggregation of keratins.

The alpha keratin helix is not a true alpha helix, as it only has 3.5 residues/turn, where the
normal alpha helix has 3.6 residues/turn. This is important for the different helices to form
tight disulfide bonds. Also, roughly every seventh residue is a leucine, so they can line up and
help the strands stick together through hydrophobic interactions.

[edit]Cornification

Cornification is the process of forming an epidermal barrier in stratified squamous epithelial

tissue. At the cellular level, cornification is characterised by:

 production of keratin

 production of small proline-rich (SPRR) proteins and transglutaminase which

eventually form a cornified cell envelope beneath the plasma membrane

 terminal differentiation

 loss of nuclei and organelles, in the final stages of cornification metabolism ceases
and the cells are almost completely filled by keratin

During the process of epithelial differentiation, cells become cornified as keratin protein is
incorporated into longer keratin intermediate filaments. Eventually
the nucleus and cytoplasmic organellesdisappear, metabolism ceases and cells undergo
a programmed death as they become fully keratinized. In many other cell types, such as cells
of the dermis, keratin filaments and other intermediate filaments function as part of
the cytoskeleton to mechanically stabilize the cell against physical stress. It does this through
connections to desmosomes, cell-cell junctional plaques, andhemidesmosomes, cell-
basement membrane adhesive structures.

Cells in the epidermis contain a structural matrix of keratin, which makes this outermost layer
of the skin almost waterproof, and along with collagen and elastin, gives skin its strength.
Rubbing and pressure cause thickening of the outer, cornified layer of the epidermis and form
protective calluses — useful for athletes and on the fingertips of musicians who play stringed
instruments. Keratinized epidermal cells are constantly shed and replaced (see dandruff).

These hard, integumentary structures are formed by intercellular cementing of fibers formed
from the dead, cornified cells generated by specialized beds deep within the skin. Hair grows
continuously and feathers moult and regenerate. The constituent proteins may
be phylogenetically homologous but differ somewhat in chemical structure and
supermolecular organization. The evolutionaryrelationships are complex and only partially
known. Multiple genes have been identified for the β-keratins in feathers, and this is probably
characteristic of all keratins.

[edit]Structural details

Keratin (high molecular weight) in bile duct cell and oval cells of horse liver

Fibrous keratin molecules supercoil to form a very stable, left-handed superhelical motif to
multimerise, forming filaments consisting of multiple copies of the keratinmonomer.[4]

Limited interior space is the reason why the triple helix of the (unrelated) structural
protein collagen, found in skin, cartilage and bone, likewise has a high percentage of glycine.
The connective tissue protein elastin also has a high percentage of both glycine and alanine.
Silk fibroin, considered a β-keratin, can have these two as 75–80% of the total, with 10–15%
serine, with the rest having bulky side groups. The chains are antiparallel, with an alternating
C → N orientation.[5] A preponderance of amino acids with small, nonreactive side groups is
characteristic for structural proteins, for which H-bonded close packing is more important
thanchemical specificity.

[edit]Disulfide bridges

In addition to intra- and intermolecular hydrogen bonds, keratins have large amounts of
the sulfur-containing amino acid cysteine, required for the disulfide bridgesthat confer
additional strength and rigidity by permanent, thermally-stable crosslinking—a role sulfur
bridges also play in vulcanized rubber. Human hair is approximately 14% cysteine. The
pungent smells of burning hair and rubber are due to the sulfur compounds formed. Extensive
disulfide bonding contributes to the insolubility of keratins, except
in dissociating or reducing agents.

The more flexible and elastic keratins of hair have fewer interchain disulfide bridges than the
keratins in mammalian fingernails, hooves and claws (homologous structures), which are
harder and more like their analogs in other vertebrate classes. Hair and other α-keratins
consist of α-helically-coiled single protein strands (with regular intra-chain H-bonding), which
are then further twisted into superhelical ropes that may be further coiled. The β-keratins of
reptiles and birds have β-pleated sheets twisted together, then stabilized and hardened by
disulfide bridges.

[edit]Filament formation

It was theorized that keratins are combined into 'hard' and 'soft,' or 'cytokeratins' and 'other
keratins'[clarification needed]. That model is now understood to be correct. A new nuclear addition in
2006 to describe keratins takes this into account.[6]

Keratin filaments are intermediate filaments. Like all intermediate filaments, keratin proteins
form filamentous polymers in a series of assembly steps beginning with dimerization; dimers
assemble into tetramers and octamers and eventually, the current hypothesis holds, into unit-
length-filaments (ULF) capable of annealing end-to-end into long filaments.

[edit]Pairing

A (neutral-basic) B (acidic) Occurrence

keratin 1, keratin 2 keratin 9, keratin 10 stratum corneum, keratinocytes

keratin 3 keratin 12 cornea

keratin 4 keratin 13 stratified epithelium

keratin 5 keratin 14, keratin 15 stratified epithelium

keratin 6 keratin 16, keratin 17 squamous epithelium

keratin 7 keratin 19 ductal epithelia

keratin 8 keratin 18, keratin 20 simple epithelium

Proline (abbreviated as Pro or P) is an α-amino acid, one of the twenty DNA-encoded amino
acids. Its codons are CCU, CCC, CCA, and CCG. It is not anessential amino acid, which
means that the human body can synthesize it. It is unique among the 20 protein-forming
amino acids in that the α-amino group is secondary. The more common L form
has S stereochemistry.
Biosynthesis
Proline is biosynthetically derived from the amino acid L-glutamate and its immediate
precursor is the imino acid (S)-1-pyrroline-5-carboxylate (P5C). Enzymes involved in a typical
biosynthesis include:[2]
 1. Glutamate 5-kinase , Glutamate 1-kinase (ATP-dependent)

2. Glutamate dehydrogenase (requires NADH or NADPH)

3. Pyrroline-5-carboxylate reductase (requires NADH or NADPH)

Zwitterionic structure of both proline enantiomers: (S)-proline (left) and (R)-proline

[edit]Structural properties
The distinctive cyclic structure of proline's side chain locks its φ backbone dihedral angle at
approximately −75°, giving proline an exceptional conformational rigidity compared to other
amino acids. Hence, proline loses less conformational entropy upon folding, which may
account for its higher prevalence in the proteins of thermophilic organisms. Proline acts as a
structural disruptor in the middle of regular secondary structure elements such as alpha
helices and beta sheets; however, proline is commonly found as the first residue of an alpha
helix and also in the edge strands of beta sheets. Proline is also commonly found in turns,
which may account for the curious fact that proline is usually solvent-exposed, despite having
a completely aliphatic side chain. Because proline lacks a hydrogen on the amide group, it
cannot act as a hydrogen bond donor, only as a hydrogen bond acceptor.

Multiple prolines and/or hydroxyprolines in a row can create a polyproline helix, the
predominant secondary structure in collagen. The hydroxylation of proline by prolyl
hydroxylase (or other additions of electron-withdrawing substituents such as fluorine)
increases the conformational stability of collagensignificantly. Hence, the hydroxylation of
proline is a critical biochemical process for maintaining the connective tissue of higher
organisms. Severe diseases such as scurvy can result from defects in this hydroxylation, e.g.,
mutations in the enzyme prolyl hydroxylase or lack of the necessary ascorbate (vitamin
C) cofactor.

Sequences of proline and 2-aminoisobutyric acid (Aib) also form a helical turn structure.[citation
needed]

In 2006, scientists at ASU discovered that solutions of TiO2 illuminated

with ultraviolet radiation can serve as an extremely cost-effective and accurate protein
cleavage catalyst. The TiO2 catalyst preferentially and rapidly cleaves protein at sites where
proline is present, while taking much longer to degrade the protein from its endpoints.[3]
Peptide bond formation with incoming Pro-tRNAPro is considerably slower than with any other
tRNAs, which is a general feature of N-alkylamino acids.[4]

[edit]Cis-trans isomerization
Peptide bonds to proline, and to other N-substituted amino acids (such as sarcosine), are
able to populate both the cis and trans isomers. Most peptide bonds overwhelmingly adopt
the trans isomer (typically 99.9% under unstrained conditions), chiefly because the amide
hydrogen (trans isomer) offers less steric repulsion to the preceding Cα atom than does the
following Cα atom (cis isomer). By contrast, the cis and trans isomers of the X-Pro peptide
bond (where X represents any amino acid) both experience steric clashes with the
neighboring substitution and are nearly equal energetically. Hence, the fraction of X-Pro
peptide bonds in the cis isomer under unstrained conditions ranges from 10-40%; the fraction
depends slightly on the preceding amino acid, with aromatic residues favoring the cis isomer
slightly.

From a kinetic standpoint, cis-trans proline isomerization is a very slow process that can
impede the progress of protein folding by trapping one or more proline residues crucial for
folding in the non-native isomer, especially when the native protein requires the cis isomer.
This is because proline residues are exclusively synthesized in the ribosome as
the trans isomer form. All organisms possess prolyl isomerase enzymes to catalyze this
isomerization, and some bacteria have specialized prolyl isomerases associated with the
ribosome. However, not all prolines are essential for folding, and protein folding may proceed
at a normal rate despite having non-native conformers of many X-Pro peptide bonds.

[edit]Uses

Proline and its derivatives are often used as asymmetric catalysts in organic reactions.
The CBS reduction and proline catalysed aldol condensation are prominent examples.

L-Proline is an osmoprotectant and therefore is used in many pharmaceutical,

biotechnological applications.

In brewing, proteins rich in proline combine with polyphenols to produce haze (turbidity).[5]

[edit]Specialities

Proline is one of the two amino acids that do not follow along with the typical Ramachandran
plot, along with glycine. Due to the ring formation connected to the Beta-carbon, the ψ and φ
angles about the peptide bond have less allowable degrees of rotation. As a result it is often
found in "turns" of proteins as its free entropy (ΔS) is not as comparatively large to other
amino acids and thus in a folded form vs. unfolded form, the change in entropy is less.
Furthermore, proline is rarely found in α and β structures as it would reduce the stability of
such structures, due to the fact that its side chain α-N can only form one hydrogen bond.
Additionally, proline is the only amino acid that does not form a blue/purple colour when
developed by spraying with ninhydrin for uses in chromatography. Proline, instead, produces
an orange/yellow colour.

[edit]History

Hermann Emil Fischer discovered proline between 1899 and 1908.

[edit]See also
 Prolinol

 Collagen

 Polyproline helix

 Peptide bond (For more discussion of cis-trans isomerization)

 Hyperprolinemia
[edit]Synthesis

The depicted scheme is for racemic proline and was taken from Vogel Practical Organic
Chemistry 5th

edition.

Cysteine (abbreviated as Cys or C)[2] is an α-amino acid with the chemical

formula HO2CCH(NH2)CH2SH. It is a non-essential amino acid, which means that it
is biosynthesized in humans. Its codons are UGU and UGC. The side chain on cysteine
is thiol, which is nonpolar and thus cysteine is usually classified as a hydrophobic amino acid.
[citation needed]
The thiol side chain often participates in enzymatic reactions, serving as
a nucleophile. The thiol is susceptible to oxidization to give the disulfide derivative cystine,
which serves an important structural role in many proteins. Cysteine is named after cystine.
Dietary sources

Although classified as a non-essential amino acid, in rare cases, cysteine may be essential
for infants, the elderly, and individuals with certain metabolic disease or who suffer
from malabsorption syndromes. Cysteine can usually be synthesized by the human body
under normal physiological conditions if a sufficient quantity of methionine is available.
Cysteine is catabolized in the gastrointestinal tract and blood plasma[citation needed]. In
contrast, cystine travels safely through the GI tract and blood plasma and is promptly reduced
to the two cysteine molecules upon cell entry[citation needed].

Cysteine is found in most high-protein foods, including:

 Animal sources: pork, sausage meat, chicken, turkey, duck, luncheon

meat, eggs, milk, whey protein, ricotta, cottage cheese, yogurt

 Plant sources: red peppers, garlic, onions, broccoli, brussels

sprouts, oats, granola, wheat germ

As other amino acids, cysteine has an amphoteric character.

(R)-Cysteine (left) and (S)-Cysteine (right) in zwitterionic form at neutral pH

[edit]Industrial sources

L-Cysteine was once obtained industrially by hydrolysis of hair and keratin. The main
contemporary route involves fermentation utilizing a mutant of E. coli. Wacker
Chemie introduced a route from substitutedthiazolines.[3] Following this technology, L-cysteine
is produced by the hydrolysis of racemic 2-amino-Δ2-thiazoline-4-carboxylic acid
using Pseudomonas thiazolinophilum.[4]
[edit]Biosynthesis

Cysteine synthesis.Cystathionine beta synthasecatalyzes the upper reaction and cystathionine gamma-
lyase catalyzes the lower reaction.

In animals, biosynthesis begins with the amino acid serine. The sulfur is derived
from methionine, which is converted to homocysteine through the intermediate S-
adenosylmethionine. Cystathionine beta-synthase then combines homocysteine and serine to
form the asymmetrical thioethercystathionine. The enzyme cystathionine gamma-
lyase converts the cystathionine into cysteine and alpha-ketobutyrate. In plants and bacteria,
cysteine biosynthesis again starts from serine, which is converted to O-acetylserine by the
enzyme serine transacetylase. The enzyme O-acetylserine (thiol)-lyase, using sulfide
sources, converts this ester into cysteine, releasing acetate.[5]

[edit]Biological functions
The cysteine thiol group is nucleophilic and easily oxidized. The reactivity is enhanced when
the thiol is ionized, and cysteine residues in proteins have pKa values close to neutrality, so
are often in their reactive thiolate form in the cell.[6] Because of its high reactivity, the thiol
group of cysteine has numerous biological functions.

[edit]Precursor to the antioxidant glutathione

Due to the ability of thiols to undergo redox reactions, cysteine has antioxidant properties.
Cysteine's antioxidant properties are typically expressed in the tripeptideglutathione, which
occurs in humans as well as other organisms. The systemic availability of oral glutathione
(GSH) is negligible; so it must be biosynthesized from its constituent amino acids,
cysteine, glycine, and glutamic acid. Glutamic acid and glycine are readily available in most
Western diets, but the availability of cysteine can be the limiting substrate.[citation needed]

[edit]Disulfide bonds

Disulfide bonds play an important role in the folding and stability of some proteins, usually
proteins secreted to the extracellular medium.[7] Since most cellular compartments
are reducing environments, disulfide bonds are generally unstable in the cytosol with some
exceptions as noted below.

Figure 2: Cystine (shown here in its neutral form) is derived from two molecules of cysteine. It features a
disulfide bond.

Disulfide bonds in proteins are formed by oxidation of the thiol groups of cysteine residues.
The other sulfur-containing amino acid, methionine, cannot form disulfide bonds. More
aggressive oxidants convert cysteine to the corresponding sulfinic acid and sulfonic acid.
Cysteine residues play a valuable role by crosslinking proteins, which increases the rigidity of
proteins and also functions to confer proteolytic resistance (since protein export is a costly
process, minimizing its necessity is advantageous). Inside the cell, disulfide bridges between
cysteine residues within a polypeptide support the protein's tertiary structure. Insulin is an
example of a protein with cystine crosslinking, wherein two separate peptide chains are
connected by a pair of disulfide bonds.

Protein disulfide isomerases catalyze the proper formation of disulfide bonds; the cell
transfers dehydroascorbic acid to the endoplasmic reticulum, which oxidises the environment.
In this environment, cysteines are, in general, oxidized to cystine and are no longer functional
as a nucleophiles.

[edit]Precursor to iron-sulfur clusters

Cysteine is an important source of sulfide in human metabolism. The sulfide in iron-sulfur

clusters and in nitrogenase is extracted from cysteine, which is converted toalanine in the
process.[8]

[edit]Metal ion binding

Beyond the iron-sulfur proteins, many other metal cofactors in enzymes are bound to the
thiolate substituent of cysteinyl residues. Examples include zinc in zinc fingersand alcohol
dehydrogenase, copper in the blue copper proteins, iron in cytochrome P450, and nickel in
the [NiFe]-hydrogenases.[9] The thiol group also has a high affinityfor heavy metals, so that
proteins containing cysteine, such as metallothionein, will bind metals such as mercury, lead,
and cadmium tightly.[10]

[edit]Post-translational modifications

Aside from its oxidation to cystine, cysteine participates in numerous posttranslational

modifications. The nucleophilic thiol group allows cysteine to conjugate to other groups, e.g.,
in prenylation. Ubiquitin ligases transfer ubiquitin to its pendant, proteins, and caspases,
which engage in proteolysis in the apoptotic cycle. Inteins often function with the help of a
catalytic cysteine. These roles are typically limited to the intracellular milieu, where the
environment is reducing, and cysteine is not oxidized to cystine.

[edit]Applications

Cysteine, mainly the L-enantiomer, is a precursor in the food, pharmaceutical, and personal
care industries. One of the largest applications is the production of flavors. For example, the
reaction of cysteine with sugars in a Maillard reaction yields meat flavors.[11] L-cysteine is also
used as a processing aid for baking.[12]

In the field of personal care, cysteine is used for permanent wave applications predominantly
in Asia. Again the cysteine is used for breaking up the disulfide bonds in the hair's keratin.

Cysteine is a very popular target for site-directed labeling experiments to investigate

biomolecular structure and dynamics. Maleimides will selectively attach to cysteine using a
covalent Michael addition. Site-directed spin labeling for EPR or paramagnetic relaxation
enhanced NMR also uses cysteine extensively.

2-Mercaptoethanol (also β-mercaptoethanol, BME, 2BME or β-met) is the chemical

compound with the formula HOCH2CH2SH. It is a hybrid of ethylene glycol, HOCH2CH2OH,
and 1,2-ethanedithiol, HSCH2CH2SH. ME or βME, as it is commonly abbreviated, is used to
reduce disulfide bonds and can act as a biological antioxidant by scavenging hydroxyl
radicals (amongst others). It is widely used because the hydroxyl group confers solubility in
water and lowers the volatility. Due to its diminished vapor pressure, its odour, while
unpleasant, is less objectionable than related thiols.

2-Mercaptoethanol may be prepared by the action of hydrogen sulfide on ethylene oxide:[1]

[edit]Reactions

2-Mercaptoethanol reacts with aldehydes and ketones to give the corresponding

oxathiolanes. This makes 2-mercaptoethanol useful as a protecting group.[2]
 [edit]Applications

[edit]Reducing proteins
Some proteins can be denatured by 2-mercaptoethanol via its ability to
cleave disulfide bonds:

cysS-Scys + 2 HOCH2CH2SH → 2 cysSH + HOCH2CH2S-SCH2CH2OH

By breaking the S-S bonds, both the tertiary structure and

the quaternary structure of some proteins can be disrupted.[3] Because
of its ability to disrupt the structure of proteins, it was used in the
analysis of proteins, for instance, to ensure that a protein solution
contains monomeric protein molecules, instead of disulfide linked
dimers or higher order oligomers. However, since 2-mercaptoethanol
forms adducts with free cysteines and is somewhat more
toxic, dithiothreitol (DTT) is more generally employed especially in SDS-
PAGE. DTT is also a more powerful reducing agent with a redox
potential (at pH 7) of -0.33 V, compared to -0.26 V for 2-
mercaptoethanol.[4]

2-mercaptoethanol is often used interchangeably

with dithiothreitol (DTT) or the odorless tris(2-carboxyethyl)phosphine
(TCEP) in biological applications, although 2-mercaptoethanol is also
the least stable.[5]

The COL1A1 gene provides instructions for making part of a large molecule called type I collagen.
Collagens are a family of proteins that strengthen and support many tissues in the body, including
cartilage, bone, tendon, skin, and the white part of the eye (the sclera). Type I collagen is the most
abundant form of collagen in the human body.
The COL1A1 gene produces a component of type I collagen called the pro-α1(I) chain. Collagens begin
as procollagen molecules, which must be processed by enzymes outside the cell to remove extra
protein segments from their ends. Each rope-like procollagen molecule is made up of three chains: two
pro-α1(I) chains, which are produced from the COL1A1 gene, and one pro-α2(I) chain, which is
produced from theCOL1A2 gene.
After procollagens are processed, the resulting mature collagen molecules arrange themselves into
long, thin fibrils. Individual collagen molecules are cross-linked to one another within these fibrils. The
formation of cross-links results in very strong type I collagen fibrils, which are found in the spaces
around cells.

Osteogenesis imperfecta (OI) is a group of genetic disorders that mainly affect the
bones. The term "osteogenesis imperfecta" means imperfect bone formation. People
with this condition have bones that break easily, often from mild trauma or with no
apparent cause. Multiple fractures are common, and in severe cases, can occur even
before birth. Milder cases may involve only a few fractures over a person's lifetime.
There are at least eight recognized forms of osteogenesis imperfecta, designated type I
through type VIII. The types can be distinguished by their signs and symptoms,
although their characteristic features overlap. Type I is the mildest form of
osteogenesis imperfecta and type II is the most severe; other types of this condition
have signs and symptoms that fall somewhere between these two extremes.
Increasingly, genetic factors are used to define the different forms of osteogenesis
imperfecta.
The milder forms of osteogenesis imperfecta, including type I, are characterized by
bone fractures during childhood and adolescence that often result from minor trauma.
Fractures occur less frequently in adulthood. People with mild forms of the condition
typically have a blue or grey tint to the part of the eye that is usually white (the
sclera), and may develop hearing loss in adulthood. Affected individuals are usually
of normal or near normal height.
Other types of osteogenesis imperfecta are more severe, causing frequent bone
fractures that may begin before birth and result from little or no trauma. Additional
features of these conditions can include blue sclerae, short stature, hearing loss,
respiratory problems, and a disorder of tooth development called dentinogenesis
imperfecta. The most severe forms of osteogenesis imperfecta, particularly type II,
can include an abnormally small, fragile rib cage and underdeveloped lungs. Infants
with these abnormalities have life-threatening problems with breathing and often die
shortly after birth.
How common is osteogenesis imperfecta?
This condition affects an estimated 6 to 7 per 100,000 people worldwide. Types I and
IV are the most common forms of osteogenesis imperfecta, affecting 4 to 5 per
100,000 people.
What genes are related to osteogenesis imperfecta?
Mutations in the COL1A1, COL1A2, CRTAP, and LEPRE1 genes cause osteogenesis
imperfecta.
Mutations in the COL1A1 and COL1A2 genes are responsible for more than 90 percent of
all cases of osteogenesis imperfecta. These genes provide instructions for making
proteins that are used to assemble type I collagen. This type of collagen is the most
abundant protein in bone, skin, and other connective tissues that provide structure and
strength to the body.
Most of the mutations that cause osteogenesis imperfecta type I occur in
the COL1A1 gene. These genetic changes reduce the amount of type I collagen
produced in the body, which causes bones to be brittle and to fracture easily. The
mutations responsible for most cases of osteogenesis imperfecta types II, III, and IV
occur in either the COL1A1 or COL1A2 gene. These mutations typically alter the
structure of type I collagen molecules. A defect in the structure of type I collagen
weakens connective tissues, particularly bone, resulting in the characteristic features
of osteogenesis imperfecta.
Mutations in the CRTAP and LEPRE1 genes are responsible for rare, often severe cases
of osteogenesis imperfecta. Cases caused by CRTAP mutations are usually classified as
type VII; when LEPRE1mutations underlie the condition, it is classified as type VIII.
The proteins produced from these genes work together to process collagen into its
mature form. Mutations in either gene disrupt the normal folding, assembly, and
secretion of collagen molecules. These defects weaken connective tissues, leading to
severe bone abnormalities and problems with growth.
In cases of osteogenesis imperfecta without identified mutations in
the COL1A1, COL1A2, CRTAP, orLEPRE1 gene, the cause of the disorder is unknown.
These cases include osteogenesis imperfecta types V and VI. Researchers are working
to identify additional genes that may be responsible for these conditions.
Read more about the COL1A1, COL1A2, CRTAP, and LEPRE1 genes.
How do people inherit osteogenesis imperfecta?
Most cases of osteogenesis imperfecta have an autosomal dominant pattern of
inheritance, which means one copy of the altered gene in each cell is sufficient to
cause the condition. Many people with type I or type IV osteogenesis imperfecta
inherit a mutation from a parent who has the disorder. Most infants with more severe
forms of osteogenesis imperfecta (such as type II and type III) have no history of the
condition in their family. In these infants, the condition is caused by new (sporadic)
mutations in theCOL1A1 or COL1A2 gene.
Less commonly, osteogenesis imperfecta has an autosomal recessive pattern of
inheritance. Autosomal recessive inheritance means two copies of the gene in each
cell are altered. The parents of a child with an autosomal recessive disorder typically
are not affected, but each carry one copy of the altered gene. Some cases of
osteogenesis imperfecta type III are autosomal recessive; these cases usually result
from mutations in genes other than COL1A1 and COL1A2. When osteogenesis
imperfecta is caused by mutations in theCRTAP or LEPRE1 gene, the condition also has
an autosomal recessive pattern of inheritance.

Osteogenesis imperfecta
Brittle bone disease
Last reviewed: August 7, 2009.
Osteogenesis imperfecta is a condition causing extremely fragile bones.
Causes, incidence, and risk factors
Osteogenesis imperfecta (OI) is a congenital disease, meaning it is present at birth. It is frequently
caused by defect in the gene that produces type 1 collagen, an important building block of bone. There
are many different defects that can affect this gene. The severity of OI depends on the specific gene
defect.
OI is an autosomal dominant disease. That means if you have one copy of the gene, you will
have the disease. Most cases of OI are inherited from a parent, although some cases are the result of
new genetic mutations.
A person with OI has a 50% chance of passing on the gene and the disease to their children.
Symptoms
All people with OI have weak bones, which makes them susceptible to fractures. Persons with OI are
usually below average height (short stature). However, the severity of the disease varies greatly.
The classic symptoms include:
Blue tint to the whites of their eyes (blue sclera)
Multiple bone fractures
Early hearing loss (deafness)
Because type I collagen is also found in ligaments, persons with OI often have loose joints
(hypermobility) and flat feet. Some types of OI also lead to the development of poor teeth.
Symptoms of more severe forms of OI may include:
Bowed legs and arms
Kyphosis
Scoliosis (S-curve spine)
Signs and tests
OI is usually suspected in children whose bones break with very little force. A physical examination
may show that the whites of their eyes have a blue tint.
A definitive diagnosis may be made using a skin punch biopsy. Family members may be given a
DNA blood test.
If there is a family history of OI, chorionic villus sampling may be done during pregnancy to
determine if the baby has the condition. However, because so many different mutations can cause OI,
some forms cannot be diagnosed with a genetic test.
The severe form of type II osteogenesis imperfecta can be seen on ultrasound when the fetus is as
young as 16 weeks.
Treatment
There is not yet a cure for this disease. However, specific therapies can reduce the pain and
complications associated with OI.
Bisphosphonates are drugs that have been used to treat osteoporosis. They have proven to be very
valuable in the treatment of OI symptoms, particularly in children. These drugs can increase the
strength and density of bone in persons with OI. They have been shown to greatly reduce bone pain and
fracture rate (especially in the bones of the spine).
Low impact exercises such as swimming keep muscles strong and help maintain strong bones. Such
exercise can be very beneficial for persons with OI and should be encouraged.
In more severe cases, surgery to place metal rods into the long bones of the legs may be considered to
strength the bone and reduce the risk of fracture. Bracing can also be helpful for some people.
Reconstructive surgery may be needed to correct any deformities. Such treatment is important because
deformities (such as bowed legs or a spinal problem) can significantly affect a person's ability to move
or walk.
Regardless of treatment, fractures will occur. Most fractures heal quickly. Time in a cast should be
limited since bone loss (disuse osteoporosis) may occur when you do not use a part of your body for a
period of time.
Many children with OI develop body image problems as they enter their teenage years. A social worker
or psychologist can help them adapt to life with OI.
Expectations (prognosis)
How well a person does depends on the type of OI they have.
Type I, or mild OI, is the most common form. Persons with this type can live a normal lifespan.
Type II is a severe form that is usually leads to death in the first year of life.
Type III is also called severe OI. Persons with this type have many fractures starting very early in life
and can have severe bone deformities. Many become wheelchair bound and usually have a somewhat
shortened life expectancy.
Type IV, or moderately severe OI, is similar to type I, although persons with type IV often need braces
or crutches to walk. Life expectancy is normal or near normal.
There are other types of OI, but they occur very infrequently and most are considered subtypes of the
moderately severe form (type IV).
Complications
Complications are largely based on the type of OI present. They are often directly related to the
problems with weak bones and multiple fractures.
Complications may include:
Hearing loss (common in type I and type III)
Heart failure (type II)
Respiratory problems and pneumonias due to chest wall deformities
Spinal cord or brain stem problems
Permanent deformity
Prevention
Genetic counseling is recommended for couples considering pregnancy if there is
a personal or family history of this condition.

Homeostasis is the maintenance of a constant internal environment within the body.

It includes

· Control of the water balance of the blood

· Control of blood sugar level

· Control of body temperature

· Control of blood urea level

Each of these internal ‘Controls’ is maintained by a separate ‘Mechanism’

All ‘mechanisms’ for homeostasis share common features:

· A specific sensor is able to detect the value of the factor which is being monitored

· Any deviation from the desired value (norm) is corrected so that the norm is more-or-less
maintained

· The corrective mechanism involves negative feedback

Negative Feedback
Control mechanism in which a change from the norm triggers off a response, or responses,
which allow the norm to be reset

BLOOD SUGAR REGULATION

The sugar carried in the blood is glucose. The level of glucose in the blood is closely
monitored and has a norm of approximately 80mg per 100cm3 blood

The regulation of glucose involves the pancreas and the liver

· In the pancreas there are groups of special cells known as ‘Islets of Langerhans’. These
cells secrete 2 hormones Insulin and Glucagon.
· If the blood sugar level rises, e.g. after a heavy meal, these cells detect this and release
more insulin and less glucagon.

· The insulin travels to the liver and ‘tells’ it to do a number of things (1) convert glucose to
glycogen(stored in the liver and muscles), (2) Convert glucose to fat

· As a result the blood sugar level falls

TEMPERATURE REGULATION

Significant variation in the internal temperature could have damaging effects on the body’s
enzymes

Humans are ‘Endotherms(Warm blooded- keep constant body temperature in hot or cold
climate)’ as opposed to ‘Ectotherms(Reptiles and Fish, their body temperatures vary
according to outside temperature)

Heat can be gained or lost from the body in four ways:

· Radiation-Transfer of heat to or from the body to other objects (via the air)

· Conduction-Transfer of heat to or from the body by direct contact with another object e.g.
radiator, window

· Convection- Transfer of heat to or from the body via moving air

· Evaporation- The loss of heat used to change water from a liquid to a vapour, e.g. when
water leaves the skin surface or gas exchange system

RESPONSE TO HEAT GAINS

· Sweating

· Flattening of hairs on the skin

· Vasodilation and dilation of shunt vessels

· More blood in the circulation-blood which is stored in the liver and spleen is released into
the circulation. More blood therefore reaches the skin surface, therefore more heat is lost

· Metabolic rate falls. This is a long-term effect under the control of the hormone thyroxine

· Lack of shivering- this spasmodic contraction of voluntary muscle to produce heat does
not occur

The response to heat LOSSES is the reverse of these changes.

ROLE OF THE BRAIN IN TEMPERATURE REGULATION

The hypothalamus in the brain acts as a thermostat and is sensitive to changes in blood
temperature. If the temperature of the blood entering the hypothalamus falls, it sends
impulses to organs causing then to reduce heat loss. The reverse happens if the temperature
of the blood entering the hypothalamus rises.

ROLE OF THE TEMPERATURE RECEPTORS IN THE SKIN

These sense changes in the external temperature and are useful in:

· Informing the brain of these changes so that the brain can alter behaviour e.g. if the
person feels a cold draught he/she may close a window

· Protection, e.g. if a person picks up an object which is very hot then he/she drops it very
quick
The synthesis of collagen, for which vitamin C is essential, proceeds in the body as one of its major
manufacturing enterprises. A person who is dying of scurvy stops making this substance, and his body
falls apart -- his joints fail, because he can no longer keep the cartilage and tendons strongs, his blood
vessels break open, his gums ulcerate and his teeth fall out, his immune system deteriorates, and he
dies.
Collagen is a protein, one of the thousands of different kinds of proteins in the human body. Most
proteins occur in only small amounts: the various enzymes, for example, are so powerful in their ability
to cause specific chemical reactions to take place rapidly that only a gram or two or even a few
milligrams may be needed in the body. There are a few exceptions. There is a great amount of
hemoglobin in red blood cells. There is even more collagen in the skin, bones, teeth, blood vessels, eye,
heart, and, in fact, essentially all parts of the body. Collagen as strong white fibers, stronger than steel
wire of the same weight, and as yellow elastic networks (called elastin), usually together with
macropolysaccharides, constitutes the connective tissue that holds our bodies together.

Like other proteins, collagen consists of polypeptide chains; the long chains of this fibrous molecule
contain about one thousand amino-acid residues, about sixteen thousand atoms. It differs from almost
all other proteins in being substantially composed of but two amino acids, glycine and hydroxyproline.
Collagen is a kind of supermolecule, however, in its three-dimensional architecture. The polypeptide
chains of the two amino acids, alternating with one another and punctuated by the presence of certain
other amino acids, are coiled in a left-handed helix. Three of these helical strands are twisted around on
another, like strands of a rope, in a right handed superhelix, to compose the complete molecule.

Understandably, the synthesis of this structure proceeds in steps. While it has been known for half a
century (these words written in 1985) that vitamin C is essential to the manufacture of collagen, the
process is only now yielding to inquiry. It appears that vitamin C is involved at every step.

First, a three dimensional stranded structure is assembled, with the amino acids glycine and proline as
its principal components. This is not yet collagen but its precursor, procollagen. A recent study shows
that vitamin C must have an important role in its synthesis. Prolonged exposure of cultures of human
connective-tissue cells to ascorbate induced an eight-fold increase in the synthesis of collagen with no
increase in the rate of synthesis of other proteins (Murad et al., 1981). Since the production of
procollagen must precede the production of collagen, vitamin C must have a role in this step -- the
formation of the polypeptide chains of procollagen -- along with its better understood role int he
conversion of procollagen to collagen.

The conversion involves a reaction that substitutes a hydroxyl group, OH, for a hydrogen atom, H, in
the proline residues at certain points in the polypeptide chains, converting those residues to
hydroxyproline. This hydroxylation reaction secures the chains in the triple helix of collagen. The
hydroxylation, next, of the residues of the amino acid lysine, transforming them to hydroxylysine, is
then needed to permit the cross-linking of the triple helices into the fibers and networks of the tissues.
These hydroxylation reactions are catalyzed by two different enzymes: prolyl-4-hydroxylase and lysyl-
hydroxylase. Vitamin C also serves with them in inducing these reactions. It has recently been shown
by Myllyla and his colleagues that, in this service, one molecule of vitamin C is destroyed for each H
replaced by OH [Myllyla et al., "Ascorbate is Consumed Stoichiometrically in the Uncoupled
Reactions Catalyzed by Prolyl-4-Hydroxylase and Lysyl Hydroxylase. Journal of Biological Chemistry
259:5403-5405. 1984]

We have come upon the two big reasons why we require for good health so much larger amounts of
vitamin C than are present in the plants we use as food. First, there is the bodies continuing need for the
synthesis of large amounts of collagen for growth and for replacement of the collagen degraded by
daily wear and tear. Second, vitamin C, in the critical reactions that assemble collagen in the tissues,
does not serve merely as a catalyst but is destroyed."