Membranes/Bio-membranes

(Definition, Types, Structural Organization and Basic Functions, Composition

and Antigenicity of Biological Membrane Components).
The study of cells has been a challenging, active area of research since the seventeenth century when Robert
Hooke first observed them through his compound microscope. Cells consist of cytoplasm, which contains
biomolecules such as carbohydrates, nucleic acids and proteins, and is enclosed by a plasma membrane. This
membrane plays an essential role in cellular protection as well as in the control and the transport of ions, nutrients
and a variety of small molecules through protein channels or processes like diffusion and endocytosis.
One of the key developments towards determining the structure of the plasma membrane was the proposal of the
“Fluid Mosaic Model” by SJ Singer and GL Nicolson in 1972. The model states that membranes are composed
of a phospholipid bilayer backbone with embedded cholesterol, proteins, and carbohydrates that gives the
membrane its fluid character. This composite structure allows the membrane to perform multiple functions such
as molecular recognition, enzymatic catalysis, cellular adhesion and membrane fusion. The most important
evolution of this model happened in 1997 with the works of Simons et al. and of Brown et al. who proposed that
membrane lipids were organized into phase-separated micro-domains, called lipid rafts, with a different local
composition and a molecular dynamic from the surrounding liquid crystalline phase. The validity of this
hypothesis was a hotly debated topic for more than a decade but has since been demonstrated by a lot of
researchers and has led to the development of technologies for detecting lateral heterogeneity in biological
membranes.

Overview
A biological membrane or bio-membrane is an enclosing or separating membrane that acts as a selectively
permeable barrier within living things.
All cells are surrounded by Biological Membranes, which all have the same basic structure. Some organelles
found in Eukaryotic Cells also have membranes. Among their roles, membranes are essential to: maintaining
the integrity of the cell and the various membrane-bound organelles within the cell by separating the
cell/organelle contents from their environment; regulating the transport of materials into and out of the cell.
Starch molecules for example, may not be able to leave the cell; responding to external and internal stimuli, cell-
to-cell cell communication and recognition, and in holding some components of metabolic reactions in place.
Biological membranes, in the form of cell membranes, often consist of a phospholipid bilayer with embedded,
integral and peripheral proteins used in communication and transportation of chemicals and ions. Bulk lipid in
membrane provides a fluid matrix for proteins to rotate and laterally diffuse for physiological functioning.
Proteins are adapted to high membrane fluidity environment of lipid bilayer with the presence of an annular lipid
shell, consisting of lipid molecules bound tightly to surface of integral membrane proteins.

The Phospholipid Bilayer

All Biological Membranes are made of the same basic structure. This is composed of molecules called
Phospholipids, which form a Phospholipid Bilayer.
Phospholipids are fats. They are composed of two Fatty Acid 'tails' and a Phosphate 'head'. The Phosphate
'heads' are Hydrophilic whereas the Fatty Acid 'tails' are Hydrophobic, meaning that Phospholipids are
Amphipathic or Amphiphilic molecules.
When placed in water, the 'heads' orientate themselves towards water molecules and the 'tails' away, meaning
that phospholipids will form a layer above water if left. If Phospholipid Molecules are completely surrounded by
water, they may form a Bilayer.
A Phospholipid Bilayer consists of two layers of Phospholipids where the 'tails' point inwards and the 'heads'
point outwards, towards water. One layer is like a mirror image of the other.
The Phospholipid Molecules are not bonded together, however, their Amphipathic Nature gives the Bilayer a
degree of stability, since the Hydrophilic 'head' cannot easily through the Hydrophobic region created by the
'tails'. The molecules can however move freely as a fluid in the plane of the Bilayer.
Small, non-polar molecules can pass through the Phospholipid Bilayer since they can 'squeeze' between the
Phospholipid Molecules and are not repelled by the hydrophobic region. Water molecules can also move
through the Bilayer, despite being polar.

At the end of the lesson, students should be able to;

1. Define bio-membrane
2. List & discuss constituents of bio-membrane, composition & distribution of lipids in the bilayer
3. Discuss the functions of bio-membrane and properties of lipid bilayer
4. Discuss diffusion of bio-membranes lipids
5. Discuss how lipid fluidity is affected by lipid composition and temperature
6. Discuss the types of proteins associated with the lipid bilayer
7. Discuss the roles of integral proteins
8. Discuss artificial membrane and its applications
9. Discuss membrane fusion
10. Explain the interaction of toxins with bio-membranes
11. Describe defense mechanisms in parasites

Definition, Types and Functions of Membrane.

What are membranes?
Membranes or bio-membranes are complex molecules composed of proteins, lipids and carbohydrate molecules.
They are sheet-like enclosed structures consisting of an asymmetric lipid bilayer with distinct inner and outer
surfaces. These sheet-like structures are formed spontaneously in water due to the amphipathic nature of lipids.
The membranes contain numerous proteins that carry out specific functions. They are structures that define and
control the composition of the space that they enclose.
Alternately, bio-membranes are lipid-protein-sugar “sheets”, in which the permeability barrier and structural
integrity are provided by the lipids; specific functions are carried out by proteins and distinctive “appearance” is
provided by sugars.

Structure of a Biological Membrane

Functions of Bio-membranes
Biological membranes vary in their precise composition, however, there are general activities and properties
common to all membranes. They play four general roles in the cell
1. The membrane surrounding the cell (plasma membrane), defines the boundary of the cell and acts as a
permeability barrier that restricts the movement of substances in or out of the cell. In a eukaryotic cell, in addition
to the plasma membrane, there also are membranes that define organelles such as the mitochondria and nucleus.
Their membranes also act as permeability barriers, so the contents of the organelles cannot mix freely with the
contents of the cytoplasm.
2. Membranes organize and compartmentalize specific activities within or around the cell. This is accomplished
by their association with proteins, which have distinct activities with different organelle membranes or distinct
regions of the plasma membrane. For example, there are specific enzymes embedded in the endoplasmic reticulum
(ER) membrane that modify proteins by the addition of a polysaccharide.
3. Membranes regulate the transport of molecules in and out of the cell, and between organelles and the cytoplasm.
This activity is regulated by specific proteins embedded in the membrane, which allow selective movement of
ions, glucose, and other small molecules.
4. The plasma membrane receives signals from the environment, including other cells. In most cases, these are
extracellular signals in the form of small molecules or proteins that are detected by specific receptors embedded
in the membrane, and result in a change in the cell. This process of receiving a chemical signal and transmitting
it to the cell is referred to as signal transduction.
Other functions include:
1. They keep toxic substances out of the cell
2. They contain receptors and channels that allow specific molecules, such as ions, nutrients, wastes, and
metabolic products that mediate cellular and extracellular activities to pass between organelles and between the
cell and the outside environment
3. They separate vital but incompatible metabolic processes conducted within organelles.

Structural Organization and Composition of Bio-membranes.

The thickness of biological membranes varies from approximately 2 to 10 nanometers depending on the cell type,
function and species. Regardless of their source, all bio-membranes have the same basic Lipid bilayer structure
and composition, however, each type of cellular membrane also has certain distinctive activities determined
largely by the unique set of proteins associated with that membrane. Lipid bilayers are sheet-like assemblies of
thousands of amphiphilic lipid molecules held together by hydrophobic interactions between their acyl chains.
Membrane lipids can be divided into three groups based on their chemical structure: glycerol-based lipids
(phospholipids), ceramide-based sphingolipids, and sterols (cholesterols). The two basic categories of membrane
proteins are integral proteins (all or part of which penetrate or span the phospholipid bilayer), and peripheral
proteins (which do not interact with the hydrophobic core of the bilayer).

Membrane Lipids
The major lipids in mammalian membranes are; Phospholipids, Glycosphingolipids and Sterol
(Cholesterol)

The Phospholipids
There are two major classes of phospholipids; Phosphoglycerides and Sphingophospholipids.
Phospholipids are unique molecules in that one end of the molecule is polar or hydrophilic with the other end
being nonpolar or hydrophobic. This type of molecule is referred to as amphiphilic or amphipathic that is having
both hydrophilic and hydrophobic regions. The nonpolar end of the lipid is composed of two fatty acids
chemically bond to either a glycerol or serine molecule. The fatty acids generally range from 14 to 24 carbon
atoms in length. One of the fatty acids is saturated whereas the other one is unsaturated with at least one cis-
double bond. Chemically bound to the glycerol or serine portion of the molecule is a single phosphate group to
which the polar end of the phospholipid is attached. These polar ends can be ethanolamine, choline, serine, or to
a lesser extent inositol. Four very common phospholipids found in membranes include: phosphatidylcholine,
phosphatidylserine, phosphatidylethanolamine, and sphingomyelin (found mainly in plasma membranes). The
sphingomyelin has its fatty acids chemically bound to a serine molecule rather than glycerol. Sphingomyelin
contains sphingosine, an amino alcohol with a long unsaturated hydrocarbon chain. In sphingomyelin, the
terminal hydroxyl group of sphingosine is esterified to phosphocholine, so its hydrophilic head is similar to that
of phosphatidylcholine. The molecules are aligned in such a manner that the inner part of the membrane is
composed of the hydrophobic ends of each fatty acid while the hydrophilic portion of each molecule either faces
the exterior or interior portions of the cell. It should be noted that the arrangement of the phospholipids within the
bilayer structure is not uniform. This asymmetric organization is due to differences in the chemical compositions
of the inner and outer monolayer portions of the phospholipid bilayer. In addition to phospholipids, the membrane
bilayer may also contain glycolipids, proteins, as well as cholesterol.

Phosphoglycerides
They are the more common class of phospholipid and consist of a glycerol backbone to which two fatty acids are
attached in ester and a phosphorylated alcohol. The fatty acid constituents are usually even-numbered carbon
molecules, most commonly containing 16 or 18 carbons. They are unbranched and can be saturated or unsaturated
with one or more cis double bonds. The simplest phosphoglyceride is phosphatidic acid

Sphingophospholipids
This is the second major class of phospholipids. Sphingophospholipids contain the sphingosine backbone rather
than glycerol. Sphingomyelin is prominent in myelin sheaths. A fatty acid is attached by an amide linkage to the
amino group of sphingosine forming a ceramide. The primary hydroxyl group (-OH) of sphingosine is esterified
to phosphorylcholine forming sphingomyelin.

Sterols (Cholesterol)
Cholesterol and its derivatives constitute another important class of membrane lipids, the steroids. It is the most
common sterol in mammalian membranes. It is found in the plasma membranes of mammalian cells and to a
lesser extent in certain organelles such as mitochondria, golgi complexes and also in nuclear membranes but is
absent from most prokaryotic cells. The basic structure of steroids is the four-ring hydrocarbon. Cholesterol has
a hydroxyl substituent on one ring. Although cholesterol is almost entirely hydrocarbon in composition, it is
amphipathic because its hydroxyl group can interact with water. Cholesterol intercalates among the phospholipids
of the membrane with its hydroxyl group at the aqueous interface and the remainder of the molecule within the
leaflet. Saturated fatty acids have straight tails, whereas unsaturated fatty acids, which generally exist in the cis
form in membranes, make kinked tails. As more kinks are inserted in the tails, the membrane becomes less tightly
packed and therefore more fluid. As much as 30 to 50 percent of the lipids in plant plasma membranes consists
of cholesterol and certain steroids unique to plants.

Lipid bilayer: This is a thin bimolecular sheet of mainly phospholipid molecules that form the structural basis
for all cell membranes. The amphipathic nature of phospholipids suggests that the two regions of the molecule
(hydrophilic head and hydrophobic tail) have incompatible solubility.

Membrane Carbohydrates
Carbohydrates found in many membranes are covalently bound either to proteins as constituents of
glycoproteins or to lipids as constituents of glycolipids. Bound carbohydrates increase the hydrophilic character
of lipids and proteins and help to stabilize the conformations of many membrane proteins. The simplest glycolipid,
glucosylcerebroside, contains a single glucose unit attached to a ceramide.

Membrane Proteins
Proteins are the major functional components in the composition of membranes and forms between 40%-80% of
the membrane depending on the source. For example, the inner mitochondrial membrane is 76% protein and 24%
is phospholipid. The plasma membranes from human red blood cell contain 44% proteins and 43% phospholipids.
The myelin from nerve fibers contains about 18% proteins with 76% phospholipids. These membrane proteins
function as receptors, enzymes, pumps, channels, structural components, antigens and as transport proteins. They
are vital components of the membrane lipid bilayer. The hydrophilic nature of peptide bond is minimized by the
helical structure of proteins. Their hydrophilic regions protrude at the inside and outside faces of the membrane
but connected by a hydrophobic region that traverses the hydrophobic core of the membrane bilayer. There are
two types of Membrane proteins: Integral proteins and Peripheral proteins. The integral proteins are those
proteins which are tightly bound to the membrane whereas the peripheral proteins are loosely bound to only one
side of the lipid bilayer. The manner in which a protein is associated with the membrane is indicative of its role.
Transmembrane proteins are integral membrane proteins since they traverse the membrane from one side to the
other.

The Integral Proteins

They constitute a high percentage of membrane protein, interact extensively with the phospholipids and require
the use of detergents for their solubilization. They span the membrane bilayer .They are asymmetrically
distributed across the membrane bilayer. They are usually globular and are amphipathic.

The Peripheral Proteins

They are bound to the hydrophilic regions of specific integral proteins. They do not directly interact with the
hydrophobic cores of the phospholipids in the bilayer.

Biological Membranes and the Transport of Molecules

One of the integral roles attributed to membranes is the regulation of materials into and out of the cell and/or
membrane-bound organelles. Due to the amphiphilic nature of the phospholipids, the ability to traverse a
membrane is a function of both the size and polarity of the molecule. Whereas very small nonpolar molecules are
able to diffuse fairly rapidly across membranes, polar molecules experience a certain degree of difficulty.
Uncharged polar molecules less than 90Da in size are capable of diffusing across membranes, whereas those
greater than 90Da in size do not. One dalton is equivalent to 1.66 x 10-24g or 1g/mol. Ions, however, are incapable
of diffusing across the membrane regardless of their size. From this it is apparent that the movement of substances
across biological membranes involves a number of different mechanisms. The major cellular transport
mechanisms are passive transport and active transport.

Passive Transport
The process whereby substances move from one side of a membrane to the other without the expenditure of
energy is referred to as passive transport or diffusion. In this type of transport, the movement of an uncharged
substance across the membrane is based solely on differences in concentration. This concentration gradient
determines the direction of transport which is from an area of greater concentration to one of lesser concentration.

Active Transport
The transport of substances against a concentration gradient is known as active transport and requires both a
carrier protein and an energy source. These transmembrane carrier proteins have been classified as uniport,
symport, and antiport carrier proteins. The uniport carriers transport a single molecule from one side of the
membrane to the other. Both symport and antiport carrier proteins involve the dependent transfer of two molecules
and as such they are also referred to as co-transporters. Symport carriers transport two molecules in the same
direction, such as amino acids and Na+. Antiport carriers, on the otherhand, transport the two molecules in
opposite directions, as in the case of Na+ and K+. The transport of macromolecules such as proteins and
polysaccharides rely on vesicular transport. In this type of transport, membranous vesicles are formed around the
substance or substances that are to be transported into or out of the cell or membrane bound organelle. Where
endocytosis refers to vesicular transport into the cell, exocytosis is the vesicular transport of substances out of the
cell. Two forms of endocytosis are: pinocytosis, and phagocytosis. The two are classified on the basis of the size
of the transport vesicles. Pinocytosis involves vesicles up to 150 nm in diameter whereas in phagocytosis the
vesicles formed are in excess of 250 nm in diameter.
The energy required for the active transport of molecules across a membrane can come from the hydrolysis of
adenosine triphosphate (ATP) or from specific ion gradients. Active transport can be classified into primary or
secondary active transport. The energy released from the hydrolysis of ATP drives the primary active transport
mechanism, whereas ion gradients drive the secondary active transport mechanisms.
Membranes are a Mosaic of Lipids and Proteins
The major components of a biological membrane are lipids and proteins. All membranes are organized as a lipid
bilayer, with proteins embedded in, or associated with, the bilayer. Carbohydrates are also present, although
much less abundant, and are found as sugars linked to either lipids or proteins in the membrane.

Membrane as a Fluid Mosaic/Two Dimensional Fluid

A membrane is not a static sheet of molecules locked rigidly in one place. Membrane molecules are held together
primarily by weak hydrophobic interaction. Most lipids and some membrane proteins are constantly in lateral
motion. Biological membrane is a two dimensional liquid/fluid of oriented lipids and globular proteins, since
membrane lipids and proteins (are constantly in lateral motion), can move pass each other along the membrane.
This movement is very rapid and can occur about 10 7 times per second. Individual lipid molecule can also rotate
very rapidly along their head-to-tail axes and flexible tails can “wave about” at their ends. Because such
movements are lateral, waving or rotational, it is referred to as lateral, waving or rotational diffusion/motion.
In contrast to lateral diffusion, lipid molecules rarely move from the monolayer they occupy to the opposite
monolayer since the lipid composition of the two layers are often different. The transfer of a phospholipid
molecule from one layer to the other is known as transverse diffusion/flip-flop.
However, in membranes of the endoplasmic reticulum where phospholipids are synthesized, there is rapid flip-
flop of lipids across the bilayer. This is achieved by proteins called phospholipid translocators or flippases.

Membrane Fluidity
Membrane fluidity is the ability of membrane to take on a more liquid-like or fluid arrangement.
Membrane fluidity is determined by several factors such as temperature, the length of the fatty acid chains,
the level of saturation of the fatty acid chains (presence of double bonds), and the presence of cholesterol.
The temperature at which lipid bilayers melts is called the transition temperature of that phospholipid. For most
bio-membranes the temperature range from 10-40oC. This is the temperature below which the lipid bilayer will
get to gel state (freeze) and above which it will become more fluid (melt or liquid state). Shorter hydrocarbon
chains and more double bonds increase membrane fluidity by decreasing the transition temperature. Cholesterol
tends to make membranes less fluid at higher temperatures, but paradoxically, high concentrations of cholesterol
make membranes more fluid at lower temperatures. Basically, membrane fluidity is a function of temperature and
lipid composition.

Temperature
The hydrophobic chains of the fatty chains can be aligned or ordered to provide a rather stiff structure in a bilayer
lipid. Increase in temperature causes the hydrophobic side chains to undergo a transition from ordered state to a
fluid arrangement. The temperature at which the structure undergoes the transition from the ordered state to
disordered state is called the transition temperature (Tm).

Lipid Composition
The multiple (unsaturated) bonds that exist in the cis configuration increase the fluidity of a bilayer by decreasing
the compactness of the side chain. Cholesterol reduces the fluidity of membranes. Transition temperature is
however indistinguishable at a high cholesterol phospholipid ratio.
The fluidity of a membrane significantly affects its functions. As membrane fluidity increases, so does its
permeability to water and other small hydrophilic molecules. The lateral mobility of integral proteins increases
as the fluidity of the membrane increases.

Diffusion (Mobility) of Membrane Lipids

Flexibility/Mobility is a notable feature of biomembranes. Membrane flexibility is their ability to change shape
without losing their integrity and becoming leaky. It is a peculiar quality of biomembranes.
Membrane flexibility is made possible by the non-covalent interactions among the lipid bilayers and the motions
allowed to individual lipids. Sterols reduce membrane flexibility. The rigid planar structure of the steroid nucleus
inserted between fatty acyl side chains reduces the freedom of neighboring fatty acyl chains to move by rotation
about their fully extended conformation.
Mobility (diffusion) of a given membrane components depends on:
a). the size of the molecule
b). its interactions with other molecules
c). temperature
d). lipid composition (tails, cholesterol)

Types of Diffusion of Membrane Lipids

1. Flip-flop diffusion/ transbilayer: This is uncatalyzed transverse diffusion
2. Catalysed transverse diffusion
3. Uncatalysed lateral diffusion

Flip-flop Diffusion
Flip-flop occurs slowly within a temperature range of 20-40oC. It requires that a polar/charged head group leave
its aqueous environment and move into the hydrophobic interior of the bilayer.
This type of movement becomes necessary during the synthesis of the bacterial plasma membrane, phospholipids
are produced on the inside surface of the membrane and must undergo flip-flop diffusion to enter the outer leaflet
of the bilayer. The flippases facilitates flip flop diffusion this provides transmembrane path that is energetically
more favourable and much faster than the uncatalyzed movement.

Different Cell Type →Different Composition of Lipids

Major Lipid Components of Selected Bio-membranes (Composition in mol %)

Artificial Membrane and Membrane Fusion

Artificial membranes are synthetically made membranes usually for separation purpose. They are produced from
organic compounds i.e. cellulose nitrate or acetate, inorganic compounds i.e. alumina etc. Separation depends on
factors such as the chemical properties, physical properties, nature of separated particles and choice of driving
force

Application of Artificial Membrane

1. Dehydrogenation of natural gas
2. Removal of microorganisms from products (dairy product)
3. Water purification-reverse osmosis,
4. Microfilteration: Membrane pore size (0.1 - 10? m) Process of separating material of colloidal size.
Ultrafilteration. Membrane pore size (0.1 - 0.01? m)
5. Dialysis
Advantages of Artificial Membranes
1. The possibility of varying the lipid content of the membranes. This allows systematic assessment of the effects
of varying lipid composition on certain function
2. Purified membrane proteins/enzymes can be integrated into these vesicles in order to assess the required factors
for the proteins to reconstitute
3. The environment of these systems can be rigidly controlled and systematically varied (e.g. ion concentrations,
ligands)

Liposomes
They are small artificial vesicles of spherical shape that can be created from cholesterol and natural non-toxic
phospholipids.
Their usefulness in drug delivery is as a result of its size, hydrophobic and hydrophilic characters. Liposomes can
also be made to entrap certain compounds inside themselves.
Examples are drugs and isolated genes. Liposomes can be used to distribute drugs to certain tissues. If certain
antibodies to certain cell surface molecules could be incorporated into liposomes so that they would be targeted
to specific tissues or tumors, the therapeutic impact would be considerable. DNA entrapped inside liposomes
appears to be less sensitive to attack by nucleases.

Membrane Fusion
This is the coming together of two membranes without loss of continuity. Mechanisms involving membrane
fusion are seen in exocytosis (release of neurotransmitters), endocytosis, cell division fusion of egg and sperm
cells and entry of membrane-enveloped virus into its host cell

Requirements for Membrane Fusion

1. They must recognize each other
2. Their bilayer structures become locally disrupted resulting in fusion of the outer leaflet of each membrane
(hemifusion)
3. Their surfaces become closely apposed which requires the removal of water molecules normally associated
with the polar head groups of lipids
4. The fusion process is triggered at the appropriate time or in response to specific signal
5. Their bilayers fuse to form a single continuous bilayer

Examples of Membrane Fusion

1. The entry of an enveloped virus (influenza virus) into a host cell
2. The release of neurotransmitters
The entry of an influenza virus into a host cell
The influenza virus is enveloped by a membrane containing hemagglutination (HA) protein. The virus enters the
host cell by inducing endocytosis at a pH of about 5. The low pH causes a conformational change in the HA
which exposes a sequence within the HA protein, this enables the protein penetrate the endosomal membrane.
Thus, the endosomal membrane and the viral membrane are connected through the HA protein. The HA protein
bends at its middle forming a hairpin shape, bringing its two ends together, this pulls the two membranes into
close apposition, causes fusion of the viral membrane and the endosomal membrane
The release of neurotransmitters
Neurotransmitters are released at synapses when intracellular vesicles loaded with neurotransmitter fuse with the
plasma membrane. This process involves a family of proteins called SNARES.

Types of SNARES
1. v-SNAREs: SNAREs in the cytoplasmic face of the intracellular vesicles
2. t-SNAREs: those in the target membranes with which the vesicles fuse
3. Other proteins involved: SNAP25 and NSF
During fusion, v- and t- SNAREs bind together and undergo a structural change that produces a bundle of long
thin rods made up of helices from both SNARES and two helices from SNAP25.
This structural change pulls the two membranes into contact and initiates the fusion of their lipid bilayers.

Techniques for Analyzing lipid and Protein Composition

The lipid composition of any given membrane can be determined by thin-layer chromatography (TLC). This
technique separates lipids in a mixture based on their differential affinity for a stationary phase and a moving
phase.
The mobility of lipids within a biological membrane can be measured by fluorescence recovery after photo-
bleaching (FRAP).
The organization of protein complexes in the lipid bilayer can be visualized by freeze-fracture electron
microscopy. The most common way of disrupting the lipid bilayer and solubilizing the proteins is through the
use of detergents such as sodium dodecyl sulfate (SDS). Once membrane proteins are isolated from the lipid
bilayer, they can be analyzed in a variety of ways.
The most widely used technique for analyzing protein composition is SDS polyacrylamide gel electrophoresis
(SDS PAGE). SDS PAGE separates proteins based on their molecular weights.

Study Questions
• Discuss the organization of phospholipids and integral proteins in all biological membranes
• Outline the functions of proteins, carbohydrate and lipids in biological membranes.
• Outline the functions of the plasma membranes in prokaryotic and eukaryotic cells.
• Compare and contrast the chemical composition of biological membranes from a variety of prokaryotic
and eukaryotic cells.
• Describe the mechanisms by which materials are transported across membranes.
• How does Temperature and Composition of membrane affect its Fluidity?
• Does the nucleus allow molecules to pass across its double membrane?

Membrane Asymmetry and Movement (Diffusion, Rotation). Membrane

Fluidity. Introduction to Receptor Function.
Biological Membranes and the Transport of Molecules
One of the integral roles attributed to membranes is the regulation of materials into and out of the cell and/or
membrane-bound organelles. Due to the amphiphilic nature of the phospholipids, the ability to traverse a
membrane is a function of both the size and polarity of the molecule. Whereas very small nonpolar molecules are
able to diffuse fairly rapidly across membranes, polar molecules experience a certain degree of difficulty.
Uncharged polar molecules less than 90Da in size are capable of diffusing across membranes, whereas those
greater than 90Da in size do not. One dalton is equivalent to 1.66 x 10-24g or 1g/mol. Ions, however, are incapable
of diffusing across the membrane regardless of their size. From this it is apparent that the movement of substances
across biological membranes involves a number of different mechanisms. The major cellular transport
mechanisms are passive transport and active transport.

At the end of the lesson, students should be able to:

1. Discuss the transport types in membranes
2. Discuss transporters

Communication through Bio-membranes

Types of Transport in Membranes
Two types of transport process occur across the membrane.
1. Non-mediated transport
2. Mediated transport

Non-mediated transport occurs through the simple diffusion process and the driving force for the transport of a
substance through a medium depends on its chemical potential gradient. Whereas mediated transport requires
specific carrier proteins. Thus, the substance diffuses in the direction that eliminates its concentration gradient; at
a rate proportional to the magnitude of this gradient and also depends on its solubility in the membrane’s non-
polar core. Mediated transport is classified into two categories depending on the thermodynamics of the system:

1. Passive-mediated transport, or facilitated diffusion also referred to as Passive (Diffusion); Simple or

facilitated via ion channels and transporters: In this type of process a specific molecule flows from high
concentration to low concentration.
2. Active transport: In this type of process a specific molecule is transported from low concentration to high
concentration, that is, against its concentration gradient. Such an endergonic process must be coupled to a
sufficiently exergonic process to make it favorable (ΔG <0)

1. Passive Transport (Diffusion)

The process whereby substances move from one side of a membrane to the other without the expenditure of
energy is referred to as passive transport or diffusion. In this type of transport, the movement of an uncharged
substance across the membrane is based solely on differences in concentration. This concentration gradient
determines the direction of transport which is from an area of greater concentration to one of lesser concentration.
Substances that are too large or polar diffuse across the lipid bilayer on their own through membrane proteins
called carriers, permeases, channels and transporters. Unlike active transport, this process does not involve
chemical energy. So the passive mediated transport is totally dependent upon the permeability nature of cell
membrane, which in turn, is function of organization and characteristics of membrane lipids and proteins.

Types of Passive Transport

A). Simple Diffusion
The process of the net movement of solutes from a region of high concentration to a region of low concentration
is known as diffusion/simple diffusion. The differences of concentration between the two regions are termed as
concentration gradient and the diffusion continues till the gradient vanishes. The passive flow of a solute from a
higher to a lower concentration due to random thermal movement. It depends on factors such as the thermal
agitation of the specific molecule, concentration gradient across the membrane i.e. transmembrane gradient of the
substrate and the solubility of that solute in the hydrophobic core of the membrane bilayer.

B). Facilitated Diffusion

The passive transport of a solute from a higher to a lower concentration mediated by specific protein transporter.
The process of the movement of molecules across the cell membrane through special transport proteins that are
embedded within the cellular membrane is known as facilitated diffusion or called carrier-mediated diffusion.
Many large molecules, such as glucose, are insoluble in lipids and too large to fit into the porins, therefore, it will
bind with its specific carrier proteins, and the complex will then be bonded to a receptor site and moved through
the cellular membrane. Hydrophilic molecules that cannot freely pass through the lipid bilayer membrane do so
passively by facilitated diffusion or by active transport. Facilitated diffusion is explained by the ping-pong
mechanism. In the ping state, it is exposed to high concentrations of solute and molecules of the solute bind to
specific sites on the carrier protein. Binding induces a conformational change that exposes the carrier to a lower
concentration of solute (pong state). This process is reversible and the net flux across the membrane depends upon
the concentration gradient.
The rate of entrance of solutes into cell by facilitated diffusion is determined by:
1. The Concentration gradient across the membrane
2. The Amount of carrier available
3. The Affinity of the solute-carrier interaction
4. The Rapidity of the conformational change for both the loaded and the unloaded carrier

Factors affecting Diffusion of a substance

1. Concentration gradient across the membrane. Solutes move from high to low concentration
2. The electrical potential across the membrane toward the solution that has the opposite charge. The inside of the
cell usually has a negative charge
3. The permeability coefficient of the substance for the membrane
4. The hydrostatic pressure gradient across the membrane. Increased pressure will increase the rate and force of
the collision between the molecules and the membrane
5. Temperature. Increased temperature will increase particle motion and thus increase the frequency of collision
between external particles and the membrane
6. Facilitated diffusion involves certain transporters or ion channels. A multitude transporters and channels exist
in biological membranes that route the entry of ions into and out of cells

C). Filtration
Filtration is the process of the movement of water and solute molecules across the cell membrane due to
hydrostatic pressure generated by the system. Depending on the size of the membrane pores, only solutes of a
certain size may pass through it. The membrane pores of the Bowman's capsule in the kidneys are very small, and
only albumins (smallest of the proteins) can filter through. On the other hand, the membrane pores of liver cells
are extremely large, to allow a variety of solutes to pass through and be metabolized.

D). Osmosis
Osmosis is the type of diffusion of water molecules across a semi- permeable membrane, from a solution of high
water potential to a region of low water potential. A cell with a less negative water potential will draw in water
but this depends on other factors as well such as solute potential (pressure in the cell e.g. solute molecules) and
pressure potential (external pressure e.g. cell wall).

Fig. 1: Osmosis. (a) In isotonic solution, there is equal concentration of solute on both sides, henceforth the water with
move back in forth. (b) In hypotonic solution, there are less solute molecules outside the cell, since salt sucks and water will
move inside the cell. The cell will gain water and grow larger, and finally burst. (c) In hypertonic solution, there are more
solute molecules outside the cell, which causes the water to be sucked in that direction which leads to the shrinkage of cells.
2. Active transport
This is the transport of a solute across a membrane against a concentration gradient. It requires energy (frequently
derived from the hydrolysis of ATP). They require both a carrier protein and an energy source. These
transmembrane carrier proteins are classified as uniport, symport and antiport carrier proteins. The uniport
carriers transport a single molecule from one side of the membrane to the other. Both symport and antiport carrier
proteins involve the dependent transfer of two molecules and as such are also referred to as co-transporters.
Symport carriers transport two molecules in the same direction, such as amino acids and Na +. Antiport carriers,
on the other hand, transport the two molecules in opposite directions, as in the case of Na+ and K+. The transport
of macromolecules such as proteins and polysaccharides rely on vesicular transport. In this type of transport,
membranous vesicles are formed around the substance or substances that are to be transported into or out of the
cell or membrane bound organelle. Endocytosis refers to vesicular transport into the cell while Exocytosis is the
vesicular transport of substances out of the cell. Two forms of endocytosis are: pinocytosis and phagocytosis.
The two are classified on the basis of the size of the transport vesicles. Pinocytosis involves vesicles up to 150
nm in diameter while phagocytosis involves vesicles formed in excess of 250 nm in diameter. The energy required
for the active transport of molecules across a membrane can come from the hydrolysis of adenosine triphosphate
(ATP) or from specific ion gradients. Active transport can be classified into primary or secondary active
transport. The energy released from the hydrolysis of ATP drives the primary active transport mechanism, while
ion gradients drive the secondary active transport mechanisms.

Types of Active Transport

A). Primary active transport
Primary active transport, also called direct active transport, directly uses energy to transport molecules across a
membrane. Example: Sodium-potassium pump, which helps to maintain the cell potential.

B). Secondary active transport

Secondary active transport or co-transport, also uses energy to transport molecules across a membrane; however,
in contrast to primary active transport, there is no direct coupling of ATP; instead, the electrochemical potential
difference created by pumping ions out of the cell is instrumental.

Similarities of Passive and active transport with substrate-enzyme interaction

1. There is a specific binding site for the solute
2. The carrier is saturable, so it has a maximum rate of transport
3. There is a binding constant (Km) for the solute and so the whole system has a Km
4. Structurally similar competitive inhibitors block transport.

Transporters
Are membrane proteins that speed the movement of a solute across a membrane by facilitating diffusion. They
are classified into carriers, permeases, transporters and channels.

Carriers
Catalyze transport at rates below the limits of free diffusion, bind substrates with high stereospecificity, are
saturable just like enzymes and function as monomeric proteins.

Permeases
Permeases are a class of membrane transport proteins which facilitate the diffusion of a specific molecule by
passive mediated transport. These are divided into the following types: Lactose permease, β-galactoside
permease, amino acid permeases,

Channels
They show less stereospecificity than carriers. They are usually not saturable. They generally allow
transmembrane movement at higher rates than carriers. They are oligomeric complexes of several identical
subunits. The permeability of a channel depends on the size, extent of hydration and extent of charge density on
the ion.

Transport systems
Uniport system: Moves one type of molecule bi-directionally.
Co-transport system: the transfer of one solute depends upon the stoichiometric simultaneous or sequential
transfer of another solute.
Symport system: moves two solutes in the same direction. Examples are the proton-sugar transporter in bacteria
and the Na+ -sugar transporters (for glucose and certain other sugars) and Na+ -amino acid transporters in
mammalian cells.
Antiport system: move two molecules in opposite directions (eg, Na+ in and Ca2+ out).

Ion channels
They are very selective, in most cases permitting the passage of only one type of ion (Na+, Ca2+, etc).

Properties of Ion Channels

1. They are composed of transmembrane protein subunits.
2. Most are highly selective for one ion; a few are nonselective
3. They allow impermeable ions to cross membranes at rates approaching diffusion limits.
4. They can permit ion fluxes of 106–107/s.
5. Their activities are regulated.
6. The main types are voltage-gated, ligand-gated, and mechanically gated.
7. They are usually highly conserved across species.
8. Mutations in genes encoding them can cause specific diseases.
9. Their activities are affected by certain drugs.

Ionophores
Ionophores are molecules that act as membrane shuttles for various ions. They contain hydrophilic centres that
bind specific ions and are surrounded by peripheral hydrophobic regions. This allows the molecules to dissolve
effectively in the membrane and diffuse transversely therein. Aquaporins are proteins that form water channels
in certain membranes.

Advanced Communication through Bio-membranes

At the end of the lesson, students should be able to:
1. Discuss active transport system
2. Discuss endocytosis and exocytosis

Active Transport System

This transport system requires that molecules are transported against concentration gradients.
They require energy which can come from hydrolysis of ATP, electron movement and light.
About four major classes of ATP-driven active transporters have been recognized (P, F, V and ABC transporters).
Examples of the P class is the Na+K+-ATPase and Ca2+-ATPase of muscle. The second class is referred to as F-
type (example is mt-ATP synthase. The V-type active transporters pump protons into lysosomes and other
structures. ABC transporters include Cystic fibrosis transmembrane regulator protein (CFTR protein), a chloride
channel implicated in the causation of cystic fibrosis. Multidrug resistance-1 protein (MDR-1 protein) which
pump a variety of drugs, including manyanti-cancer agents out of the cells

Plasma membrane Na+K+- ATPase

This is an important enzyme that regulates the intracellular concentration of Na+ and K+.
Usually, cells maintain a low intracellular Na+ concentration and a high intracellular K+ concentration coupled
with a net negative electrical potential inside. ATPase is the pump that maintains these ionic gradients and is
activated by Na+ and K+. It pumps Na+ out and K+ into cells. The ATPase is an integral membrane protein
containing a transmembrane domain allowing the passage of ions and cytosolic domains that couple ATP
hydrolysis to transport. It has catalytic centers for both ATP and Na+ on the cytoplasmic side of the plasma
membrane with K+ binding sites located on the extracellular side of the membrane.
The phosphorylation by ATP of three Na+-binding sites on the cytoplasmic surface of the cell induces a
conformational change in the protein leading to transfer of 3 Na+ ions from the inner to the outer side of the
plasma membrane. 2 molecules of K+ bind to sites on the protein on the external surface of the plasma membrane
resulting in de-phosphorylation of the protein and transfer of the K+ ions across the membrane to the interior.
Therefore, 3 Na+ ions are transported out for every 2 K+ ions entering. This creates a charge difference between
the intra and extracellular compartments, making the intracellular compartment more negative. ATPase is
inhibited by cardiac drugs such as oubain and digitalis. The Na+K+-ATPase can be coupled to various other
transporters such as those involved in transport of glucose.

Endocytosis
This is a process by which cells take up large molecules. It is a mechanism for regulating the content of certain
membrane components e.g. hormone receptor. It can be used to study how cell function. DNA transfection
depends on endocytosis. It is responsible for the entry of DNA into the cell.
Endocytosis requires energy (usually from ATP), Ca2+ and contractile elements in the cell (possibly the
microfilament system). Endocytotic vesicles are generated when segments of the plasma membrane invaginate,
enclosing a small volume of extracellular fluid and its contents. The vesicle pinches off when the fusion of plasma
membranes seals the neck of the vesicle at the original site of invagination. This vesicle fuses with other
membrane structures thereby ensuring the transport of its contents to other cellular compartments. Most
endocytotic vesicles fuse with primary lysosomes to form secondary lysosomes which contain hydrolytic enzymes
hence, are specialized organelles for intracellular disposal

Types of Endocytosis
1. Phargocytosis
2. Pinocytosis

Phargocytosis: Phargocytosis involves the ingestion of large particles such as viruses, bacteria, cells or debris
and granulocytes. It occurs only in specialized cells such as macrophages. Macrophages may ingest 25% of their
volume in an hour

Pinocytosis: This is a property of all cells that leads to the cellular uptake of fluid and fluid contents.

Types of Pinocytosis
1. Fluid-phase pinocytosis. It is a non-selective process in which the uptake of a solute by formation of small
vesicles is simply proportionate to its concentration in the surrounding extracellular fluid. Fibroblasts are
examples, they internalize their plasma membrane at about one third the rate of macrophages
2. Absorptive Pinocytosis. This is a receptor-mediated selective process primarily responsible for the uptake of
macromolecules for which there are fixed numbers of binding sites on the plasma membrane. The vesicles formed
are derived from invaginations that are coated on the cytoplasmic side with a filamentous material called coated
pits. Absorptive pinocytosis of extracellular glycoproteins requires that the glycoproteins carry specific
carbohydrate recognition signals. These recognition signals are bound by membrane receptor molecules which
play a role analogous to that of the LDL receptor. A galactosyl receptor on the surface of hepatocytes is helpful
in the absorptive pinocytosis of asialoglycoproteins from circulation. Moreover, acid hydrolases taken up by
absorptive pinocytosis in fibroblasts are recognized by their mannose 6-phosphate moieties. Viruses which cause
hepatitis, poliomyelitis and AIDS initiate their damage by absorptive pinocytosis. Moreover, iron toxicity also
begins with excessive uptake due to endocytosis

Exocytosis: This releases certain macromolecules from the cell to extracellular space. It is involved in membrane
remodeling when the components synthesized in the golgi apparatus are carried in vesicles to the plasma
membrane. Signal for exocytosis is usually hormone which when it binds to a cell-surface receptor, induces a
local and transient change in Ca2+ concentration. Ca2+ triggers exocytosis.

Neurotransmission
Membranes of the neurons have an asymmetry of inside-outside voltage (electrical potential).
They are electrically excitable as a result of the presence of voltage-gated channels. Hence, appropriate
stimulation by a chemical signal mediated by a specific synaptic membrane receptor, opens the channels in the
membrane to allow the rapid entry of Na+ or Ca2+ so that the voltage difference rapidly collapses and the segment
of the membrane is depolarized. The gradient is quickly restored by the action of the ion pumps present in the
membrane. When large areas of the membrane are depolarized in this manner, the electrochemical disturbance
propagates in wavelike form down the membrane, generating a nerve impulse.
Myelin sheets, formed by Schwann cells, wrapped around nerve fibers provide an electrical insulator that
surrounds most of the nerve and greatly speeds up the propagation of the wave (signal) by allowing ions to flow
in and out of the membrane only where the membrane is free of the insulation (at the nodes of Ranvier). The
myelin membrane is highly rich in lipids, this confers a tremendous insulating property on it. Demyelination and
impaired nerve conduction are features of certain diseases such as Multiple sclerosis and Guillain-Barr syndrome

Glucose transport
The entrance of glucose into the cell is the first step in energy utilization. A number of glucose transporters are
involved depending on each tissue. Glucose enters the erythrocyte by facilitated diffusion called GLUT1. In the
skeletal muscle and adipocytes, glucose enters by a specific transport system enhanced by insulin. Glucose
transport in the small intestine involves the binding of Na+ and glucose to different sites on a Na+-glucose
symporter located at the apical surface. Na+ moves into the cell down its electrochemical gradient and drag
glucose with it. Therefore, the greater the Na+ gradient the more glucose enters and if Na+ in extracellular fluid is
low, glucose transport stops. In a bid to ensure a steep Na+ gradient, this Na+-glucose symporter is dependent on
gradients generated by the Na+-K+-ATPase which maintains a low intracellular Na+ concentration. The
transcellular movement of glucose in this case involves one additional component; a glucose uniporter: which
allows the glucose accumulated within the cell to move across the basolateral membrane

Isolation of sub-cellular organelles

At the end of the lesson, students should be able to:
1. Review cellular organelles
2. Discuss the process involved in the isolation of cellular organelles

Sub-cellular organelles
Organelles are organized structures within the cell e.g. the nucleus, mitochondria, ribosomes, golgi bodies,
endoplasmic reticulum etc. They are separated by centrifugation based on the difference in their size, density,
shape

Steps in the isolation of cell organelles

1. Homogenization
2. Sub-cellular fractionation by centrifugation
3. Marker assays

Homogenization
This is the disruption of cells under conditions that prevent deterioration aimed at isolation of morphologically
intact and functionally active organelles and microsomal fractions. It is the first step in the isolation of sub-cellular
organelles. It involves breaking the cell membrane.

Common Homogenization Techniques

1. Dounce homogenization: crushing of cells between two revolving solid surfaces
2. Filteration: cells are forced through smaller pores in a filter
3. Grinding: where cells are ground by swirling with glass beads
4. Sonication: where cells are bombarded with ultrasonic vibrations
5. Solubilization: in which cell membranes are dissolved by detergents such as triton X-100
Enzyme digestion is also used to remove cell-wall constituents. The method of choice depends on the type of
tissue to be homogenized and the specific purpose of the experiment. During homogenization, isotonic sucrose is
added to the homogenization buffer to prevent osmotic rupture of organellar membranes. After homogenization,
the homogenate is spurn at low speed to remove any intact cells along with large cellular debris. This is followed
by subcellular fractionation.

Differential centrifugation
This is the movement of subcellular particle in a centrifugal field. This movement sedimentation) results from the
interaction between a particle’s weight, the resistance it encounters in moving through a suspension medium and
the relative centrifugal force exerted on the particle. Under a given centrifugal force, particles that are relatively
large or dense will sediment more rapidly than particles that are smaller and lighter. The order of sedimentation
is typically from most to least dense; nuclei, mitochondria, lysosomes, plasma membrane, endoplasmic reticulum
and contractile vacuoles. It precedes density-equilibrium centrifugation.

Density-equilibrium centrifugation
In Density-equilibrium centrifugation, subcellular organelles are layered on a density gradients and subjected to
a very high centrifugal force. The density gradient is formed by layering increasing concentration of sucrose
solutions in a centrifuge tube. Other solutions such as cesium chloride, Percoll can be used. These two solute ns
when spun spontaneously set up a density gradient, thereby alleviating the need manually layer sucrose solutions
of varied concentration.
During centrifugation, organelles initially layered on the density gradient will sediment until they arrive at the
region of the gradient where the density of the suspension is equal to their own.

Marker enzymes
Indicate the presence of an enzyme. Acid phosphatase is located in the lysosomes, succinate dehydrogennase is
located in the mitochondria. ATP synthase is present in the inner mitochondrial membrane. Galactosyl transferase
is present in the golgi apparatus. Glucose-6-phosphatase is present in microsomes. Galactosyl transferase is
present in the Golgi apparatus.
This helps to monitor where each enzyme activity is found during a cell fractionation protocol.
Marker enzymes also provide information on the biochemical purity of the fractionated organelles. The presence
of unwanted marker enzyme activity in the preparation indicates the level of contamination by other organelles.
The degree of enrichment for the desired organelle is determined by the specific activity of the target marker
enzyme. Electron microscopy is generally used as a final step to assess the preparation’s purity and the
morphology of the isolated organelle.

Study Questions
Discuss the isolation of subcellular organelles

Cell membranes and toxins

At the end of the lesson, students should be able to:
1. Discuss the mechanisms by which toxins interfere with the host’s cell membrane
2. Discuss the defense mechanisms in parasites
3. Discuss mode of action of polymyxins

Cell membranes play significant roles in the interaction of pathogenic microbes with the host cells. About 50%
of the membrane volume is made up of transmembrane, peripheral and lipid linked proteins arranged in a bilayer.
Membrane lipid is important in providing cellular signaling, membrane trafficking as well as membrane
microdomain organization. Infectious organisms rely on the multifunctional role of lipids to modulate cell
processes to ensure their survival.
Toxins
Toxins are powerful pathogenicity factor produced by macro and microorganisms which mediate drastic
interactions of the pathogens on the organism’s host. Classically, bacteria toxins are divided into endotoxins and
exotoxins. Endotoxins: membrane compounds of gram negative bacteria which elicits inflammation in the host.
Exotoxins: secreted proteins which act locally and at distance of the bacteria colonization site. The invasion of
cells by pathogens/toxins is marked by binding to carbohydrate moieties exposed by a lipid or a protein in the
plasma membrane of target cells. Cholera toxins binds with its?-subunit to the ganglioside GM1 in the intestinal
cells, Pseudomonas aeruginosa attaches to respiratory cells by binding to asialoGM1 and asialo-GM2 through
type IV pili. Influenza A virus initiates its uptake by binding to sialic acids in the host cell membranes.
Glycosphingolipids are important for self-induced endocytosis of toxins and viruses. Phosphatidylinositol-
phosphate (PIP) is also important for the uptake of pathogens. They are essential components of the cell
membranes involved in signaling events; vesicle trafficking. One strategy by which invasive bacteria manipulate
the PIP is the translocation of effector proteins, which act as phosphatidylinositol phosphatases. A second option
to interfere with the PIP metabolism is the engagement of specific host cell receptors.

Shigella Dysenteriae
It secretes two types of enterotoxins; Shigella toxins I and II. In humans they cause serious complications in the
GIT such as haemolytic colitis. The binding affinity of the?-subunit of Shiga toxin (StxB) and cholera toxin
(CtxB) to individual Gb3 and GM1 molecules respectively is very low. But the cooperative binding of multiple
lipid molecules markedly increases the apparent affinity of the toxin to its receptor. After binding, Stx is
internalized by clathrindependent as well as clathrin-independent endocytosis. Cholera toxin has been found to
be associated with caveolae and is efficiently endocytosed into cells devoid of caveolin-1

Defense mechanisms in Parasites

The interplay between parasite survival strategies and the host defense mechanisms is the basis for the relationship
that exists between the host and the infecting parasite. Parasites have devised strategies for their survival while in
the host; the parasite aims at propagating within the host and be transmitted to subsequent host, while the goal of
the host is to limit the infection. Parasites have evolved mechanisms for evading the immune system of the host.
Below are some of the mechanisms

Protozoan immune evasion strategies

Antigenic variation
In Plasmodium, different stages of the life cycle express different antigens. In trypanosomes, there is a whole
range of variant specific surface groups (VSG) which protects the underlying membranes.
As the level of antigen increases, a small fraction of the population switches to producing a coat of a new VSG
with an antigenic character circulating antibodies are no longer able to recognize

Strategy of avoidance
Some pathogens reduce antigenicity so that they are not recognized as foreign. The?2-macroglobulin is present
on the surface of adult schistosomes and has a potent anti-protein activity. If attached to a molecule with the
appropriate conformation, it may also act as a proteinase inhibitor preventing breakdown of parasite tissues.
Plasmodium lives inside Red Blood Cells (RBCs) which have no nucleus, when infected, it is not recognized by
immune cells. Other stages of Plasmodium live inside liver cells. Plasmodium ookinetes develop in serosal
membrane and are beyond reach of phagocytic cells (hemocytes). Leishmania parasites and Trypanosoma cruzi
live inside macrophages.

Enzymatic role
Enzymes are produced that are capable of inactivating reactive oxygen intermediates resulting from the host
inflammation defense systems. The antioxidant enzymes counteract oxidative burst. Some of the enzymes are
superoxide dismutase, catalase, glutathione peroxidase, glutathione S- transferase (GST). Glutathione S-
transferase is located in the external tegument of schistosomes.
Shedding or replacement of surface
Example: Entamoeba histolytica.

Immunosupression
Manipulation of the immune response e.g. Plasmodium.

Anti-immune mechanisms
Leishmania produce anti-oxidases to counter products of macrophage oxidative burst Helminth immune evasion
mechanisms in the vertebrate host.

Large size
It is difficult for immune system to eliminate large parasites. Primary response is inflammation to initiate
expulsion, often worms are not eliminated.

Coating with host proteins

Tegument of cestode and trematode worms, is able to adsorb host components e.g. RBC Ags, thus giving the
worm the immunological appearance of host tissue. Schistosomes take up host blood proteins, e.g. blood group
antigens and MHC class I & II molecules, therefore, the worms are seen as “self”.

Molecular Mimicry
Synthesis of surface proteins similar to host proteins by the parasite which are unrecognized as foreign. In
tapeworm, synthesis of antigen that resemble blood group of MHX antigen of the host. Depression of the host
immune response.

Anatomical Seclusion
Trichinella spiralis can live inside mammalian muscle cells for many years.
Shedding or replacement of surface
Examples; hookworms, trematodes

Immunosupression
Manipulation of the immune response. High burdens of nematode infection often carried with no outward sign of
infection.

Polymyxins
They are antibiotics produced by Paenibacillus polymyxa. There are two main types; polymyxin B and polymyxin
E (colistin). They are usually used in treating gram negative bacterial infections. They however have little or no
effect on gram positive bacteria. This is because the cell wall of gram positive bacteria is too thick to permit
access to the membrane. Their neuro and nephrotoxicities make them last line therapy.

Study Questions
1. What is membrane fluidity?
2. Explain the physiological roles of integral protein
3. Discuss the isolation of subcellular organelles
4. What is endocytosis and exocytosis?
5. Describe the active transport system
6. List the advantages of artificial membranes
7. What is membrane fusion? Mention two instances of membrane fusion
8. List other applications of artificial membrane
9. Distinguish between active and passive transport
10. What are the factors that affect diffusion of a substance?
11. What are transporters? What are the differences between channels and carriers?