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
• 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
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: Harper’s Illustrated
Biochemistry, 28th ed., 2009: 406-412.