Biochemistry of Membrane

27 August 2014
DR. dr. Agnes Kwenang
Department of Biochemistry
Medical Faculty
Hasanuddin University
 Membrane functions
Serve as barriers to separate contents of
cell from external environment or
contents of organelles form remainder of
the cell.

Membranes compartmentalize and

segregate intracellular events, separate
cells from one another, and segregate
organ function.
 Membranes mediate the regulation of
cellular functions by acting as selective
barrier.

Membranes localize specific enzyme

systems and provide a semisolid-phase in
an otherwise aqueous environment.
 Proteins in cell membrane have many functions
transport of substances across the membrane
enzymes that catalyze biochemical reactions
receptors on exterior surface that bind external
ligands (e.g., hormones, growth factors)
mediators that aid ligand-receptor complex in
triggering sequence of events (second messengers
that alter metabolism are produced inside the
cell)
 Plasma membranes has selective
permeabilities
Channels and pumps
for ions and substrates
Specific receptors
for signals (e.g., hormones)
Exchange materials with extracellular
environment
exocytosis and endocytosis
 Membranes form specialized
compartments
Organelles with specialized functions
e.g., mitochondria, ER, Golgi complex
Localize enzymes
Excitation-response coupling
Energy Transduction
photosynthesis, oxidative
phosphorylation
 Internal Water Is Compartmentalized
Intracellular Fluid
(2/3 of total water)
rich in K+ and Mg2+,
phosphate major anion
protein higher
Extracellular Fluid
(1/3 of total water)
high Na+ and Ca+,
chloride major anion
glucose higher
 Structure

Membranes are made of proteins, lipids,

and carbohydrates, which are attached to
either membrane proteins or lipids.

The ratio of protein to lipid is not fixed.

It can widely from 1 : 4 to 4 : 1
 Composition of membranes varies
within and between cells

Membrane lipids.
Major lipids in mammalian membranes
- Phospholipids
- Glycosphingolipids
- Cholesterol
Phospholipids - two major classes
1. phosphoglycerides are more common
* glycerol backbone
* two fatty acids in ester linkage
-usually even-numbered carbons
(C16, C18)
-unbranched, either saturated or
unsaturated, C18 or 20:4 5,8,11,14
-phosphorylated alcohol
- phosphatidic acid (1,2-diacylglycerol
3-phosphate) is simplest -- key
intermediate in formation of all other
phospholipids
 Phospholipids - two major classes
2. Sphingomyelins
sphingosine backbone (rather than glycerol)
fatty acid attached by amide linkage
primary hydroxyl group of sphingosine esterified
to phosphocholine
prominent in myelin sheaths
Glycosphingolipids
sugar-containing lipids
e.g., cerebrosides and gangliosides
.also derived from sphingosine
.differ from sphingomyelin in group
attached to primary hydroxyl
group of sphingosine
- sphingomyelin -phosphocholine
- cerebroside - single hexose (glucose
or galactose)
- ganglioside - chain of 3 or more
sugars (at least one is sialic acid)
Sterols
most common sterol
cholesterol
almost exclusively in plasma
membrane
lesser amounts in mitochondria,
Golgi, nuclear membranes
generally more abundant
toward outside of plasma
membrane
intercalates among
phospholipids of membrane
with its hydroxyl group at
aqueous interface and
remainder of molecule within
leaflet
A. Membrane lipids are all amphipathic
They have both hydrophobic and
hydrophilic regions (like
detergents)
polar head group
nonpolar tails
Saturated fatty acids - straight
tails
Unsaturated fatty acids
(generally cis) - kinked tails
 a. Membrane lipids spontaneously form
bilayers in aqueos media, burying their
hydrophobic tails and living hydrophilic
ends exposed to the water.
Amphipathic phospholipids have two
regions with incompatible solubilities
in aqueous solvent, organize into
thermodynamically favorable form.
 e.g, micelle
 Bimolecular layer (bilayer) can also satisfy
thermodynamic requirement of amphipathic molecule

only ends or edges of bilayer sheet exposed to unfavorable

environment
can eliminate by folding sheet back upon itself to form
enclosed vesicle with no edges.
Closed bilayer is essential property of membrane
impermeable to most water-soluble molecules
b. Membrane lipids undergo lateral
diffusion, that is, side-to-side or
translational movement, and flexing.
1). The lateral movement of membrane lipids is very fast.
They change places with their nearest neighbor in less
than 1 u sec.
2) The flexing motion of phospholipids is greatest at the
hydrophobic end, which opposes the polar end group.
a). This is more pronounced with saturated fatty acids
than with unsaturated, since unsaturated fatty
acids have cis-configurations at the double bonds,
which limit their ability to flex.
b). Cholesterol decreases the ability of phospholipids to
translate and flex.
c. Phospholipids do not readily flip-flop that is ,
pass from one side of the bilayer to the other.
When the phospholipids do flip-flop, it is a slow
process that takes hours.
d. Phospholipids are asymmetrically distributed in
plasma membranes.
1). Cholin phospholipids constitute the outher
face of the bilayer.
2). The amino phospholipids are found constitute
inner, or cytoplasmic, face of the bilayer.
e. Glycolipids are found in the outer face of the
bilayer with their sugar residues exposed at
the surface of the cell.
It is believed that the complex oligosaccharides
play a role in cell-to-cell interactions.
 Lipid-soluble materials
Gases (oxygen, CO2, nitrogen)
little interaction with solvents,
readily diffuse through
hydrophobic regions of
membrane
Lipid-derived molecules (e.g.,
steroid hormones)
readily transverse bilayer
Organic nonelectrolyte molecules
diffusion dependent upon oil-
water partition coefficients
(the greater lipid solubility,
the greater its diffusion rate
across membrane)
2. Membrane proteins.
Non-lipid-soluble molecules
Proteins are also amphipathic molecules
inserted into lipid bilayer
form channels for movement of ions and small
molecules
serve as transporters for larger molecules
 Side chains determine
hydrophobic nature
6 strongly hydrophobic side chains,
few weakly hydrophobic, remainder
hydrophilic
amphipathic proteins have
hydrophobic region transvering
bilayer and hydrophilic regions
protruding inside and outside of
membrane
protein content varies with membrane
enzymes, transport proteins,
receptors
What is the effect of unsaturated
fatty acids?
as more kinks added, membrane becomes
less tightly packed, more fluid
 Membranes and components are
dynamic structures
Lipids and proteins in membranes turn over
different lipids and proteins have individual
turnover rates, may vary widely
membrane may turn over more rapidly than
any of its constituents
 Membranes Are Asymmetric Structures

Irregular distribution of proteins

within membrane
External location of
carbohydrates attached
to membrane proteins
Regional asymmetries
villous border of mucosal cells
gap junctions, tight junctions,
synapses
 Phospholipid asymmetry
choline-containing phospholipids located mainly in
outer leaflet
phosphatidylcholine, sphingomyelin
aminophospholipids preferentially located in inner
layer
phosphatidylserine, phosphatidylethanolamine
cholesterol generally present in larger amounts on
the outside
Phospholipids
 Must be limited transverse mobility (flip-flop)
half-life of asymmetry in synthetic bilayers is several
weeks
enzymes for phospholipid synthesis are located on
cytoplasmic side of microsomal membranes
flippases
phospholipid exchange proteins
 Membrane proteins exhibit two basic types of
interactions with membranes.
A. Integral proteins are hard to dissociate from
membranes. They interact extensively with the
hydrocarbon tails of lipids, and they often span the
lipid bilayer.

Trans membrane proteins are integral proteins that

are exposed both on the outer face of the membrane
and on the inner, or cytoplasmic, face.
In other words, their peptide chains extend across
the width of the membrane.
Integral membrane proteins
interact with phospholipids,
require detergents for
solubilization
usually globular,
amphipathic
may span bilayer many
times
asymmetrically distributed
across bilayer
orientation determined
during insertion in
bilayer
B. Peripheral proteins are
bound by hydrogen bonding
or electrostatic interactions
to the of exposed surface
integral proteins.
do not interact directly
with phospholipids
do not require detergent
for release
weakly bound to
hydrophilic regions of
specific integral proteins
 e.g., ankyrin, bound to integral protein band 3 of
erythrocyte membrane
spectrin, a cytoskeletal structure within erythrocyte,
bound to ankyrin
plays important role in maintenance of biconcave
shape of erythrocyte
 Artificial membranes model membrane
function
Mixtures of one or more phospholipids treated (e.g.,
sonication) to form spherical vesicles liposomes
can control lipid content to examine effects of lipid
composition on certain functions
purified membrane proteins can be incorporated into
these vesicles to access factors required for function
environment can be controlled and varied (e.g., ion
concentrations)
can be made to entrap compounds inside (e.g., drugs,
isolated genes) for drug delivery, gene therapy
C. Movement of proteins in plasma membranes.
1.The orientation of proteins in membranes is very constant, and
it is highly unlikely that membrane proteins flip-flop
2. In 1972 Singer and Nicolson proposed the fluid-mosaic model
of membrane structure. This model proteins proposes that
membrane proteins, for the most part, can move laterally in
the matrix of the lipid bilayer and their rate of movement
depends on the fluidity of the membrane.
3. The fluid-mosaic model, while largely correct, is somewhat
oversimplified in that many proteins are restricted in their
lateral movement in order to fulfill specific cellular functions.

,
 4. Devices used by cells to limit protein
diffusion in the lipid bilayer include:
a. Confinement: to limited areas
b. Cell junctions
c. Increases in mass by aggregation
d. Cross-links by extrinsic elements
e. Links to cytoplasmic components of
the cytoskeleton
5. The need for glycoproteins to have their carbohydrate
moiety on the outside, in contact with the extracellular
environment, limit the movement of glycoproteins
.
 Fluid mosaic model
This model is often
to icebergs
(membrane
proteins) floating in
a sea of
predominantly
phospholipid
molecules.
- lateral diffusion
*integral proteins
and phospholipids
can move within
the plane of the
membrane.
D. Membrane fluidity
1. The fact that some membrane components can move
in the lipid bilayer of the membrane is very important for
cell function. A number of factors influence the fluidity of
membranes, which in turn influences the physiologic
function.
2. The fluidity of membranes depends largely on the nature
of the packing and interaction of the fatty acyl chains in
membrane phospholipids.
a. Long-chain saturated fatty acids pack closely and
interact , strongly producing a relatively rigid structure

(i.e., one with low fluidity)

b. With an increase in temperature, some of the trans C-C
bonds become gauche (i.e., rotated 120o) and more
fluid structure results
 c. The change in fluidity is seen as the temperature
rises above the melting temperature ( Tm ).
d. The longer the chain length of fatty acids the higher
is the Tm
e. Unsaturated fatty acids with their cis double bonds
do pack closely; this results in more fluid structures,
in which the Tm is lowered.
f. The greater the number of double bonds , the lower
is the Tm and the greater is the fluidity of the
membrane.
3. In more highly evolved animal, cholesterol reduces
membrane fluidity by preventing the movement of fatty
acyl chains.
Phase changes (fluidity) of membrane are dependent
upon lipid composition
hydrophobic chains of fatty acids can be highly ordered
rigid structure
with temperature, side chains undergo transition from
ordered state (gel-like or crystalline phase) to disordered
(liquid-like or fluid) phase
transition temperature (Tm)
longer, more saturated fatty acid chains interact more
strongly, cause higher Tm
unsaturated chains tend to fluidity, compactness
 Cholesterol modifies fluidity of membranes
At temperatures below Tm it interferes with the
interaction of hydrocarbon tails of fatty acids and
increases fluidity.
At temperatures above Tm it limits disorder because
it is more rigid than tails of fatty acids and cannot
move in membrane to same extent, thus limits
fluidity.
At high cholesterol:phospholipid ratios, transition
temperatures are abolished.
 Fluidity significantly affects membrane functions
As membrane fluidity , so does permeability to water and
other small hydrophilic molecules
Lateral mobility of integral proteins increases
If active site of integral protein resides exclusively in
hydrophilic regions, changing fluidity probably has
little effect on activity
If protein involved in transport, with transport
components span membrane, lipid phase effects may
significantly alter transport rate.
EXAMPLE: Insulin receptor - As concentration of
unsaturated fatty acids in membrane increased (grow in
unsaturated. fatty acid rich medium), fluidity increases,
receptor binds more insulin.
 Some protein-protein interactions within plane of
membrane can restrict mobility of integral proteins
 Asymmetry of proteins and lipids
maintained during membrane assembly

Fusion of a vesicle with

the plasma membrane
preserves the
orientation of any
integral proteins
embedded in the vesicle
bilayer
 Signal Sequences Target Many Proteins
Many proteins carry signals that
target them to their destination
Major sorting decision - synthesis on
free or membrane-bound
polyribosomes
-cytosolic branch
.no signal peptide, delivered to
cytosol
.can be directed to mitochondria,
nuclei, peroxisomes by specific
signals
 Signal Sequences Target Many Proteins
rough ER branch (Secretory or
exocytotic pathway)
contain signal peptide
many destined for various membranes
(ER, Golgi, lysosomes, and plasma
membrane) and for secretion
certain proteins sorted in Golgi for
delivery to lysosomes
proteins destined for secretion carried
in secretory vesicles
regulated secretion (secretory vesicles)
constitutive secretion (transport vesicles)
 Signal Hypothesis - Entry into ER

Blobel and Sabatini - explanation for difference

between free and membrane-bound ribosomes
All ribosomes have the same structure, distinction
dependent upon protein possessing signal
sequence
 Synthesis of secretory
1. N-terminal signal sequence is
proteins
synthesized
2. Signal bound by SRP, complex
docks with SRP receptor on ER
membrane
3. Signal sequence binds to
translocon, internal channel opens,
inserted into translocon
4. Polypeptide elongates, signal sequence cleaved
5. ER chaperones prevent faulty folding, carbohydrates added to specific
residues
6. Ribosomes released, recycle
7. C-terminus of protein drawn into ER lumen, translocon gate shuts, protein
assumes final conformation
References

1. Baynes JW, Dominiczak: Medical

Biochemistry, 2009:87.
2. Davidson VL, Sittman DB: Biochemistry,
3rd ed., 1994:4-7.
3. Murray RK, et al: Harpers Illustrated
Biochemistry, 28th ed., 2009: 406-412.