Biology 331 Summer 2014

Dr. Lars Petersen Part II (July 23 end)

Office: BI 442
Tel.: 403-
E-mail: lfpeters@ucalgary.ca

Dr. T. Don Nguyen Part I (start July 21)
Office: BI 387
Tel.: 403-220-8167
E-mail: nguyendt@ucalgary.ca

E-mail to make appointments

E-mails will be replied within 24 hours (48 hours on
weekends)

Grade components
Midterm exam (July 21) 35%

Tutorial quizzes 10%

Tutorial term project 20%

Final exam 35%
(sometime between August 15 and 18)
Terms you are expected to be already
familiar with
You need to know the following in terms of their:
composition
function

Ribosome Bacteria /Prokaryote
DNA & deoxyribonucleotides Eukaryote
RNA & ribonucleotides Archaea
Sugar (carbohydrate) Multicellular
Lipid Unicellular
Enzyme Transposon
Membrane Promoter
Periodic table of elements Gene = open reading frame (ORF)
Protein & amino acids acid and base chemistry

10 m
1 m
100 mm
(10 cm)
10 mm
(1 cm)
1 mm
Human height
Length of
some nerve
and muscle
cells
Chicken
egg
Frog egg
Paramecium
100 m
U
n
a
i
d
e
d

e
y
e

Plasma Membrane
(membrane lipids and proteins)
Eukaryote Prokaryote
1
0
-
1
0
0

m

0
.
1
-
1
0

m

The plasma membrane is an essential
feature for all cells
Essential roles of membrane
1) Compartmentalization
2) Biochemical scaffold
3) Selective permeability
barrier
4) Solute transport
5) Signalling
6) Intercellular interaction
7) Energy transduction
8) Flexibility
CO
2
+RuBP
PGA
2
Acid hydrolases
1
ADP+P
i
ATP

4
H
+
7
H
2
O
3
6
IP
3
Ca
2+
5
8
Membranes role # 1: Compartmentalization
Membranes are barriers between sets of
molecules such as proteins
eg. Outside of cell vs. inside Inside of an organelle
vs. cytoplasm

Provides an environment specialized for the
function / biochemical reactions that take place
within the compartment

Most prokaryotes lack internal membranes,
therefore they do not have internal organelles
Membranes role # 2: Scaffold for biochemical
activities and cytoskeleton attachment
Membranes can act as surfaces for
biochemical reactions/cellular
processes
Some reactions are more efficient in
membranes than in solution

Membranes interact with the internal
cytoskeleton; connects plasma
membrane to cytoskeleton

mb
Attachment
protein
Membranes role # 3: Selective permeability barrier
Restricts movement of ions: different
ionic compositions on opposite sides
of the membrane.

Allows gradients to be established

Different concentrations of the same
ion on opposite sides of the
membrane can form
eg. Important for nerve and muscle
function and energy
mb
H
+
H
+
H
+
H
+
H
+
H
+
H
+ H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
IN OUT
Membranes role # 3: Selective permeability barrier
Membranes role # 4: Transporting solutes
Membranes are sites where the movement of
molecules from one compartment to the other.
eg. Ion channels, transporters
Signals from one side of membrane elicit a response on the
other side; (eg. response to hormones, growth factors,
neurotransmitters)
Membranes role # 5: Response to external signals
Membranes role # 6: Intercellular interactions
Sites where neighboring cells interact
adhesion, communication/signals, metabolic
interactions
Example: sperm-egg fusion produces signals during
embryo development that prevent other competing
sperm from entering the egg
cell
cell
Conjugation Signalling
Surface receptor
Interactions
Fibers of
extracellular
matrix (ECM)
Enzymatic
activity
Phospholipid
Cholesterol
CYTOPLASM
CYTOPLASM
Cell-cell
recognition
Glycoprotein
Intercellular
junctions
Microfilaments
of cytoskeleton
ATP
Transport
Signal
transduction
Receptor
Signaling
molecule
Attachment to the cytoskeleton
and extracellular matrix (ECM)
Membranes role # 7: Energy transduction
Sites of energy conversion one form of
energy alters to another
eg. membranes of chloroplasts and mitochondria
photosynthesis and cellular respiration
eg. rhodopsin proteins in cells of the eye

Light
retina
Human eye
Light energy gives
rhodopsin the
power to transport
ions across the
membrane
Membranes role # 8: Flexibility and deformability
Changes in plasma membrane when cells
change shape
cells crawling on a substratum





Changes in membrane organization when
vesicles pinch off from plasma membrane or
internal membranes
Amoeba proteus
Membrane
deformations
during cell
division
Chemical composition of
membrane
Plasma membrane models
2013 John Wiley & Sons, Inc. All rights reserved.
1960s
1970s to date
Major components: lipids and proteins
Lipids and proteins arrange in a lipid bilayer:
Act as the structural backbone of the membrane
Major component that estabilishes a permeability
barrier


Membrane proteins: specific functions
A lipid:protein ratio varies greatly depending on membrane
type and cell type
Membrane proteins can vary between different
membranes or cell types depending on the function of the
membrane







Carbohydrates (sugars)
Glycolipids: sugars-attached lipids
Glycoproteins: sugar-attached proteins
Lipid composition affects
membrane properties and
membrane protein function
Three major types: phospholipids,
sphingolipids, and sterols.
Lipid composition is different in
different membranes
Membrane lipids
Phospholipids (phosphoglycerides) are the most abundant
membrane lipid.
Esterification of glycerol to:
- Phosphate
- Two fatty acids chains
Membrane lipid type # 1: Phospholipids
Fatty (acyl) acid chains
Are hydrophobic, unbranched hydrocarbons ~16 20
carbons long. (eg. C16 palmytate; C18 sterate)






Fatty acid tails may be:
polyunsaturated (>1 double bond)
monounsaturated (1 double bond eg. C18 oleate,
C=C)
saturated (no double bonds)
H
3
C
COOH

COOH

C
10
C
12
C
6
C
16
H
3
C
C
8
Phospholipids are diverse in
head group and backbone
structure.

Usual additions to phosphate:
Choline phosphatidylcholine
Ethanolamine
phosphatidylethanolamine
Inositol phosphatidylinosinol
Hydrophilic
Hydrophobic
Phospholipids: hydrophilic +
hydrophobic = amphipathic.
In membranes, phospholipids
are organized into lipid
bilayers with polar head
groups on each surface and
fatty acyl chains in the core
Hydrophilic
Hydrophobic
Phospholipids arrange into
a lipid bilayers that consist
of 2 leaflets

Polar head groups face
aqueous environment

Hydrocarbon tails form
hydrophobic inner core

Membrane lipid type # 2: Sphingolipids
Sphingolipids play an important role in the nervous system and
are targets for infectious disease-causing microorganisms.
Glycolipids: substituting a carbohydrate molecule onto
a ceramide.







CH
2
C C CH
OH

H
2
N

H

H

CH (CH
2
)
n
CH
3
C O

R

Non-polar
HO

+
Sugar
Ceramide
Glycolipids
Minor components of the membrane
ABO blood group antigens are glycolipids

Gal GlcNAc Gal Glu
Fuc
Gal GlcNAc Gal Glu
Fuc
Gal GlcNAc Gal Glu
Fuc
GlcNAc
Gal
O antigen
A antigen
B antigen
Membrane lipid type # 3:
Sterols and similar compounds
50% of the membrane
smaller and less amphipathic than
other membrane lipids
campesterol
(plants: phytosterols)
cholesterol
(animals: zoosterols)
ergosterol
(fungi)
sterols
diploptene
(bacteria: hopanoids)
Cholesterol inserts between phospholipids:
The small hydrophilic hydroxyl group faces the membrane
surface.
The rest is embedded in the lipid bilayer.
Its flat and rigid rings interfere with movement of
phospholipid fatty acid tails
Cholesterol decreases permeability and increases
durability of the membrane.


Cholesterols stiffen
the membrane
Sterols and similar compounds are terpenoids
(isoprenoids)
Terpenoids are very diverse in functions and structures, and
found in all living organisms.
Terpenoids are precursors for sterol and cytochrome
synthesis in eukaryotes.
Terpenoids are derived from isopentenyl pyrophosphate
(IPP) (characterized by a 5-carbon isoprene unit).

Isoprene
Isopentenyl pyrophosphate (IPP)
Membrane lipids and fluidity
Phospholipids in a bilayer:
1. Flex and bend
2. Rotate
3. Diffuse laterally in the plane of the membrane
4. Flip-flop from leaflet of the bilayer to the other leaflet
Occurs rarely on its own
Phospholipids can be found in different amounts in each leaflet of
the membrane, with aid of special proteins called flippases
SM: sphingomyelin
PC: phosphatidylcholine
PS: phosphatidylserine
PE: phosphatidylethanolamine
PI: phosphatidylinositol
Cl: cholesterol
2013 John Wiley & Sons, Inc. All rights reserved.
1. Temperature
2. Acyl chain length
3. Acyl chain saturation
4. Sterol content
Mobility of lipid bilayers is
influenced many factors
Factor # 1: Temperature
Lipid bilayers are more fluid at higher temperature due
to "melting" of lipid-lipid interactions
Temperature at which phase transition occurs is
dependent upon the lipids present in the bilayer
transition temperature

Temperature
Viscosity
Transition point: melting
temperature (T
m
)

Temperature
Viscosity
T
m
gel/solid phase
fluid/liquid phase
Factor # 2: Length of fatty acid acyl side chains
Viscosity
16-carbon fatty acid
acyl chains
12-carbon fatty acid
acyl chains
Temperature
T
m
T
m
Factor # 3: Unsaturated bonds
in fatty acid acyl chains
Saturated bonds decrease fluidity.
Unsaturated bonds increase fluidity.
Organization of saturated fatty acyl chains is more
regular than that of chains with unsaturated bonds.
Unsaturated bonds introduce "kinks" into the chains.
kink in
chain due
to double
bond
Viscosity
Temperature
16-carbon saturated
16-carbon mono-
unsaturated
T
m
T
m
Factor # 4: Sterols
- At low temperatures: Cholesterol ring structures keep fatty
acyl chains from interacting lowers membrane viscosity
- At high temperatures: Cholesterol ring structures are more
rigid than fatty acyl chains increases membrane viscosity

Membrane proteins
Membrane Proteins
Carry out various functions in the membranes
Highly variable:
The amount of protein relative to lipid changes
between cell types and between different intracellular
membranes
Proteins may not be distributed homogenously
in membranes
eg. neuromuscular synapse apical-basal
membranes of epithelial cells
Three types: integral, peripheral, and lipid-
anchored.
Membrane protein type # 1: Integral
Integral (intrinsic) proteins
pass through lipid bilayer, and
cannot be removed without
distruption of the bilayer
(detergents)

a) Single spanning
b) Multiple spanning
Transmembrane strand
Membrane protein type # 2: Peripheral
Peripheral proteins:
- Are not inserted into the
bilayer;
- Are found on either side
of the membrane;
- Interacts with
phospholipids or with
integral proteins.
- Can be removed by high
salt, low/ high pH, etc.
without disrupting bilayer
Membrane protein type # 3: Lipid-anchored
Lipid-anchored proteins do not
pass all the way through the lipid
bilayer.
- Hydrophobic amino acid
groups allow proteins to insert
into internal (cytoplasmic)
leaflet
- Glycosylphosphatidylinositol
(GPI)-anchored proteins:
covalently bonded to
phosphatidylinositol by
glycosylation.

Proteins are distributed asymmetrically across
cellular membranes
Most sugar groups of glycoproteins and glycolipids on
outer side of plasma membrane
Proteins can be mobile in lipid bilayers
Experiments showing that proteins are mobile in
membranes will be discussed in tutorials.
Cellular membranes are considered to be fluid mosaics
ie. Molecules are not homogenously distributed in
membranes and most are mobile within the plane of the
membrane.
Domains of cholesterol and sphinogomyleins can aggregate
together to form lipid rafts.