1.

Membrane Diversity &

Asymmetry

2.Red Blood Cell

 Membrane Diversity
 The ratio of lipid to protein in eukaryotic cell
membranes

 Diversity of membrane lipids

 Protein diversity

 Varies among membrane types as a result of cellular

function
 Membrane Diversity
 Myelin sheath

 Red blood cell membrane

 Mitochondrial membrane
Eg. Myelin sheath:
 Insulated wire: 79% lipid, 18% protein & 3%
carbohydrate

An electrical insulating
Layer wrapped around the
axon which increases the
speed of electric conduction
 Inner mitochondrial membrane
– 76% protein & 24 % lipid by weight

 Human erythrocyte membrane

– 49% protein, 43% lipid & 8% carbohydrate
 Membrane Asymmetry
 Inner & outer membrane leaflets of various membranous
structures have different lipid compositions
– PE and PS are usually mostly in the inner leaflet
– PC and sphingomyelin tend to be in the outer leaflet.

 Probably has to do with the

– Different curvatures of the 2 leaflets
– Sizes of the various head groups & packing forces between
them.

 Provides different physico-chemical properties

appropriate for different interactions.
 Membrane Asymmetry

 Phospholipid flip-flop: Helps maintain membrane

asymmetry

 Movement of a phospholipid within the same

leaflet is rapid and frequent

 Movement of a phospholipid from one leaflet to

the opposite leaflet is extremely slow.
Asymmetry of Membranes
 Membrane Asymmetry
 Integral proteins are inserted into membrane
asymmetrically, during biosynthesis.
– A specified topological arrangement for each protein.
– NB determined by cellular machinery responsible for
protein translocation

 Eg. Glycophorin
– N-terminus on extracellular side of the membrane
– C-terminus in the cytosol.
– Every glycophorin molecule has the same arrangement.
 Approximate Lipid Compositions of
Different Cell Membranes
PERCENTAGE OF TOTAL LIPID BY WEIGHT
LIPID LIVER CELL RED BLOOD MYELIN MITOCHONDRION ENDOPLASMIC E. COLI
PLASMA CELL PLASMA (INNER & OUTER RETICULUM BACTERIUM
MEMBRANE MEMBRANE MEMBRANES)

Cholesterol 17 23 22 3 6 0
Phoshatidlycholine 24 17 10 39 40 0

Phosphatidylethan- 7 18 15 25 17 70
olamine
Phosphatidylserine 4 7 9 2 5 Trace

Sphingomyelin 19 18 8 0 5 0

Glycolipids 7 3 28 Trace Trace 0

Others 22 13 8 21 27 30
 Red blood cell membrane
 Primary function is the transport of respiratory
gases to and from the tissues.

 To achieve this task the red cell should be capable

of traversing the microvascular system:

 Without mechanical damage

 The cell should retain a shape which facilitates gaseous

exchange.
Red blood cell
 Red cell membrane
 In order to meet with the demands of function, the red
cell membrane should be extremely tough yet highly
flexible.

 This strength and flexibility of the red cell membrane is

due to:
1. The design of its protein cytoskeleton

2. The way the cytoskeleton interacts with the membrane lipid

bilayer.
Red cell membrane
 Composition of the red cell
membrane
 The red cell membrane consists of:

–Proteins~50%

–Lipids~40%

–Carbohydrates~10%
 Composition of the red cell
membrane
CARBOHYDRATES

 They occur only on the external surface of the

red cell.

 Composed of glycoproteins and glycolipids.

 Composition of the red cell

PROTEINS

 These are either:

-Peripheral or
-Integral, penetrating the lipid bilayer.
Composition of the RBC Membrane
LIPIDS
 Lipid components of the red cell membrane
are:

– 30% free unesterified cholesterol.

– 10% glycerides and free fatty acids.

– 60% phospholipids
 Red cell membrane lipids
 All of the lipids associated with red cells are
present in the cell membrane.

 The mature red cell has no capacity to

synthesize lipids

 Alteration in membrane lipid content can only

occur by exchange with plasma lipids.
 RBCM Lipids
 The red cell membrane phospholipids are:
– Phosphatidyl choline
– Phosphatidyl ethanolamine
– Sphingomyelin
– Phosphatidylserine.

 Phospholipid molecules are characterized by a polar

head group attached to a non-polar fatty acid tail.

 Thus the phospholipids in the cell membrane tend to

arrange themselves in a bilayer
 Chemical Composition of RBC
Membrane
 SM, sphingomyelin
 PC, phosphatidylcholine
 PS, phosphatidylserine
 PE, phosphatidylethanolamine
 PI, Phosphatidylinositol
 Cl, cholesterol
 RBCM Lipids
 The membrane cholesterol is unesterified and lies
between the 2 layers of the lipid bilayer.

 Cholesterol conc - an important determinant of

membrane surface area & fluidity.

 An increase in membrane cholesterol leads to an

increased surface area and decreased deformability.
 RBCM Proteins
 The red cell peripheral proteins interact to form a
cytoskeleton.
 The cytoskeleton acts as a tough supporting framework for
the lipid bilayer.
 Four peripheral proteins play a key role in the structure of
the red cell cytoskeleton, these are:
– Spectrin, which is the most abundant and consists of two chains
a and b spectrin.
– Ankyrin
– Protein 4.1
– Actin
 Red cell membrane proteins
SPECTRIN
 The most abundant, consists of two chains,
– α and β, wound around each other to form heterodimers
which then self-associate head to head to form tetramers.

 These tetramers are linked at the tail end to actin and

are attached to protein band 4.1.

 At the head end, the b-spectrin chains attach to

ankyrin which connects to band 3(anion channel).
Protein 4.2 enhances this interaction.
 RBCM Integral Proteins
 RBC membranes have 2 abundant transmembrane
proteins
– Glycophorin forms a carbohydrate coat around the cells. Heavily
glycosylated (60%).
• Glycophorins are rich in sialic acid, which gives the red blood cells a very
hydrophilic-charged coat. This enables them to circulate without adhering
to other cells or vessel walls.
– The anion channel (band 3 protein) mediates exchange of
bicarbonate & chloride ions
 These integral membrane proteins are linked by band 4.1
protein & ankyrin to a flexible meshwork consisting of
spectrin
 The membrane skeleton enables RBCs to resist strong
shearing forces
 Red cell membrane proteins
BAND 3
 One of the integral proteins
 Has two major functions within the red cell
membrane:
– Its primary function is to facilitate anion transport
via the red cell membrane.
– It is an important binding site for cytoskeletal and
other red cell proteins.
 Red cell membrane proteins
Na+/K+ ATPase
 This enzyme catalyses the hydrolysis of ATP to
ADP, liberating energy in this process.

 Each ATP molecule hydrolysed via this system

results in the ejection of three Na+ ions from
the cell and the transport of two K+ ions into
the cell.
 Red Blood Cell Membrane
 Erythrocyte membrane has been studied
intensively due to:
– Ready availability
– Relative simplicity
• Only have 1 membrane, the plasma membrane

 Cytoplasmic contents can be released by osmotic

haemolysis to give ghosts
– Pure membranes
 Functions of red cell membrane
 To separate the contents of the cell from the
plasma.

 To maintain the characteristic shape of the red

cell.

 To regulate intracellular cation conc.

 To act as the interface between the cell and its

environment via membrane surface receptors.
 Blood Group Antigens
 Antigens found on the RCM: responsible for blood
group determination.
 Found on both the protein and the carbohydrate
components of the membrane i.e glycoproteins
and the glycolipids.
 Some antigens are considered as minor membrane
proteins.
 Others are found on larger membrane proteins.
 Some blood group antigens are absorbed passively
from the plasma, e.g. Peter, Lisa and Tendai.
 Isolation of RBCM proteins

 Integral proteins such as band 3 &

glycophorins

 Can be dissociated from the membrane only

by organic solvents & detergents such as
SDS and Triton-X 100
 Red Blood Cell Membrane
 Red blood cell membrane proteins with
carbohydrate units:
– Anion channel
– Glycophorin
 Require treatment with periodic acid-Schiff
reagent (PAS)
 PAS is a staining method used to detect
polysaccharides such as glycogen, and
mucosubstances such as glycoproteins,
glycolipids and mucins in tissues.