Council of Scientific & Industrial Research

VOLUME – 4

CELLULAR ORGANIZATION, CELL

COMMUNICATATION & CELL SIGNALING
 CONTENT
1. Biomembrane 1

2. Transport Across A Cell Membrane 7

3. Protein Targeting 29

4. Cell Signaling 68

5. Cell Cycle 101

6. Cancer 128

7. Apoptosis 141

8. Cytoskeleton 152

9. Cell Adhesion and Communication 166

 BIOMEMBRANE
All cells and cell organelles are made up of a basic structure that is called as
biomembrane.eg. Endoplasmic reticulum and Golgi apparatus. All membranes
consist of three substances: - Lipids, Proteins, Carbohydrate.

The ratio of lipids and proteins depends on the function of cell. Every cell has a
different combination of protein, lipid and carbohydrate as follows:

The biomembrane is made up of phospholipids. It is impermeable for water-soluble

molecules, ions and biological molecules. Membranes are formed by a lipid bilayer
of phospholipids. Phospholipids are made up of a polar heads (water) and fatty acid
tails (oil). Polar heads are hydrophilic and tails are hydrophobic.

Natural phospholipids fatty acids have one or more double bonds. It causes kink
formation in the interior of the membrane and make the hydrocarbon chains
difficult to be packed together. The polar head region is variable in charge and fatty
acids (R) chains also vary in length from Cię to Czas well as in their degree of
unsaturation. i.e. double bond.

Biomembrane Components:
Biomembrane has mainly three components.
(1) Proteins (2) Lipid (3) Carbohydrate

Proteins
All biomembranes consist of proteins, glycolipids steroids and cholesterol. Due to
the presence of different types of proteins in biomembranes, the cell may vary in
its functions. Membrane receptor proteins transfer the signals between the cells
internal and external environments. Transport proteins involve in transport of
molecules across the membrane. Many types of proteins are present in membrane.
Some are attached to the surface of membrane and many proteins are embedded
in the membrane, that's why the cell membrane is not smooth, it seems to be rough.
Membrane enzymes may play a crucial roles in many cellular activities like
oxidoreductase, transferase or hydrolase. Cell adhesion molecules gives a platform

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to the cells to identify and interact with each other. For eg. Protein involved in
immune response. There are mainly four types of proteins, found in the
biomembrane.
(A) Intrinsic or Integral protein
(B) Extrinsic or Peripheral protein
(C) Protein anchor proteins
(D) Lipid anchor proteins

Integral Proteins:
These proteins are embedded in the membrane. These are attached permanently
in a bio membrane. These can be separated only by using nonpolar solvents,
denaturing agents and detergents. Integral proteins can be single pass, double pass
and multi-pass proteins. The protein which crosses single time a membrane and
having a-helix structure is called a single pass. If a proteins a-helix structure passes
twice in the membrane, it is called a double pass integral proteins and if a protein
passes multiple time in the membrane, it is termed as multi-pass intrinsic protein.
 𝑯𝒆𝒍𝒊𝒙 𝒃𝒖𝒏𝒅𝒍𝒆 𝒑𝒓𝒐𝒕𝒆𝒊𝒏𝒔: Present in all types of biomembranes
 𝜷-Barrel proteins (Porins): Porins are beta-barrel proteins that cross a cellular
membrane and act as a pore, through which molecules can diffuse. They are
large enough to allow passive diffusion. i.e. they act as channels that are specific
to different types of molecules. Porins are present in the other membrane of
gram negative bacteria such as E.Coli, mitochondria, chloroplast and some gram-
positive bacteria of the group Mycoleta.

E.Coli has double membrane, the cell wall (outer membrane) & plasma-membrane
(inner membrane). The outer membrane are more permeable than inner due to this
porins channels, which form aqueous channels through the lipid bilayer. The outer
membrane protect the intestinal bacterium from harmful agents (eg-antibiotics,
bile salts & proteases) but permits the uptake and disposal of small hydrophilic
molecules including nutrients and waste products. It also allows passage of
disaccharides and other small molecules as well as phosphate.

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Integral monotopic proteins:
These proteins attach to only one side of biomembrane and do not span the whole
way across.
Other examples of integral proteins are glycophorins and band 3 proteins.
(A)Glycophorin: - It is one of the example of integral proteins in red blood cells. It is
rich in sialic acid. This acid gives a very hydrophilic charged coat to RBC. This
makes them to circulate without disturbing the other cells. These are distributed
in 5 types. Glycophorin A, Glycophorin B, Glycophorin Cand Glycophorin E.
(B) Band - 3 - It is 14 transmembrane protein. This gene protein is encoded by the
SLC4A1 (Solute carrier family 4 member Gene in humans. This is called as Band 3
anion transport protein or anion exchanger 1 (AE1). Many activities like protein
anchor, transporter activity, protein and ankyrin binding, inorganic anion
exchanger activity, regulation of intracellular pH, chloride transport, sodium
transport, anion transport are occured by this protein.
(C) Peripheral membrane proteins or extrinsic proteins: These proteins are
temporarily attached to the biomembrane or to the integral proteins by a non-
covalent, electrostatic or hydrophobic interactions. These can be dissociated by
a polar reagent treatment. Integral and peripheral proteins may be post-
translationally modified, with the addition of glycosyl phosphotidylinositol (GPI)
prenyl chains and fatty acid. Which may be anchored in the lipid bilayer.
Examples of these proteins are ankyrin and band 4.1.
(i) Ankyrin: Ankyrins are the mediator protein between spectrin and integral
protein band 3. Thus it links between the plasma membrane and
cytoskeleton. These have binding sites for b-subunit of spectrin and at least
12 families of integral membrane protein. This linkage plays a very important
role in the integrity of plasma membrane, ion channels, ion exchangers and
ion transporters in the plasma membrane.
(ii) Band 4.1: This protein is associated with the cytoskeleton and encoded by the
EPB41 gene in the human. This protein plays a very crucial role in maintaining
the structure and skeleton of erythrocytes.

Protein anchor proteins:-

These type of proteins are involved in Apoptosis. These are water-soluble proteins
but these can aggregate and associate irreversibly with the lipid bilayer and become
reversibly or irreversibly membrane associated.

Lipid-anchor proteins:
LAP is also known as lipid-linked proteins. They are located on the surface of the cell
membrane. They are covalently attached to lipids. These proteins are embedded

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within the cell membrane. These proteins are linked to a certain fatty acid like
myristate and palmitate. These anchors the protein to the cell membrane this
protein to the cell membrane. These play an important role in protein functions.
Lipids The involvement of phospholipids, cholesterol and proteins occurs in the
fluidity of the membrane. Lipid packaging and its components influence the fluidity
of the membrane. The fluidity of the membrane is crucial for membrane functions.
The membrane behaves as two-dimensional fluids.

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Membrane Fluidity
The ratio of saturated and unsaturated fatty acids determines the fluidity in the
membrane at cold temperature. Factors affecting the fluidity of biomembrane;

(a) Temperature: At low and high temperatures cholesterol increases and

decreases the fluidity of membrane respectively. When fluidity increases, it
prevent membrane lipids packing close together. Temperature decides what
can enter or leave the cell. Cell functions it's best at normal physiological
temperature, which is 98.6 degrees Fahrenheit in warm-blooded animals like
the human.
(b) Lipid Composition: If saturated fatty acids are compressed by decreasing
temperatures, they press in on each other, making a dense rigid membrane. If
unsaturated fatty acids are composed, the kinks push adjacent phospholipid
molecules away, which helps maintain fluidity in the membrane.
(c) The different form of fatty acid: Transform of fatty acid is more packed than the
cis form of fatty acid. Thus trans fatty acid is solid and cis fatty acid is liquid.

(d) Cholesterol: Cholesterol serves as a buffer, preventing lower temperatures from

inhibiting fluidity and preventing higher temperatures from increasing fluidity.
Cholesterol helps to separate the phospholipids in our cell plasma membranes
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 at the high concentrations so that the chains of fatty acids cannot come together
and crystalize. So, cholesterol prevents extremes whether too fluid or too firm
in the consistency of the cell membrane.

Lipid Rafts:
Some microdomains of cholesterol and sphingolipids from discrete membrane are
known as lipid rafts. These clusters move laterally within the plasma membrane and
get associated with specific membrane proteins. Two types of lipid rafts are (1)
planar (Non-coveolar) (2) Caveolae. Caveolae are flask-shaped invagination of the
plasma membrane that contains caveolin protein. Caveolin is an intrinsic
cholesterol binding membrane protein. Although the function of lipid rafts remain
to be fully understood, they play an important role in processes such as cell
movement (Cholesterol trafficking) and the uptake of extracellular molecules by
endocytosis as well as in cell signalling. The lipid composition of a bilayer also
influences its thickness which in turn may play a role in localizing proteins to a
particular membrane. It is found that sphingomyelin associates into a more gel-like
& thicker bilayer than phospholipids do. Similarly, cholesterol and other molecules
that decrease membrane fluidity increase membrane thickness. Because
Sphingomyelin tails are already optimally stabilized, the extra addition of
cholesterol has no effect on the thickness of a sphingomyelin bilayer. Lipid rafts are
the centres of organizing for the assembly of signalling molecules, influencing
membrane fluidity and membrane protein trafficking and regulating
neurotransmission and receptor trafficking lipid rafts are more ordered and tightly
packed than it's surrounding structure of biomembrane. Lipid rafts are present in
plasma membranes and other parts of cells like Lysosomes and Golgi complex.

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TRANSPORT ACROSS A CELL MEMBRANE
The transport processes are critically important to all living cells because all living
cells necessarily exchange the substances with their surrounding environment like
nutrients, wastes, toxicant and the inorganic electrolytes are also able to pass
through a living membrane.

Osmosis
Osmosis is the diffusion of water (solvent) molecules from a more dilute solution
(high concentration of water) to more concentrated solution(less concentration of
water) down the water potential through a semi-permeable membrane. This is a
vital physical process found among all biological systems. Osmosis is a spontaneous
process.

The pressure required to maintain the equilibrium where the net movement of
solvent is zero is known as osmotic pressure. Osmotic pressure depends directly on
the molar concentration of the solute. Osmoregulation is done by osmosis process.
By osmoregulation organism can homeostat water and salt amounts in the body.

All type of cells maintaining a concentration gradient of all these various

metabolites across their functional membranes. Such concentration gradients are
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maintained by the membranes as a huge amount of potential energy. For example,
Nat and K ions gradient across the membranes regulates the brain function, nerve
impulses transmission, muscle contractions and all other normal functions of heart,
kidney, liver, stomach and such other organs.

The cell membrane is a bilayer of phospholipids that is nearly impermeable to the

water-water soluble molecules and ions. But these substances are transported
across the membrane at a high rate to fulfil the physiological and metabolic needs
of a cell. This critical problem is solved with the help of some membrane's integral
proteins that work as transporters. The transporting materials either diffuse
through the channel formed by the proteins or they may be carried out or in by the
carrier proteins.

The membrane transport processes are of two types based on according to the
thermodynamics and the kinetics.
1. Passive transport
2. Active transport

Passive transport
Transport of ions or molecules from high concentration to low concentration
without energy expenditure. Now LET'S TALK about why molecules move from high
concentration to low concentration. The molecules move because of their own
kinetic energy (K.E.=3/2 KT, where T is absolute Temperature). High concentration
and high temperature allow more collision between the molecules.

Passive transport is also of two types, the first one is the simple diffusion or the
second one is the facilitated diffusion.

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Simple diffusion: Simple diffusion is the transport of ions or molecules from high
concentration to low concentration without energy expenditure across a
membrane of hydrophobic phospholipids and cholesterol bilayer.

This is the simplest transport process that has no need of any protein. Gases like 𝑂2
𝑁2 and 𝐶𝑂2 and the small uncharged molecules like urea, ethanol can easily move
by simple diffusion through the plasma membrane. Because the transportation of
such uncharged molecules across the membrane is an entropic process, in which
the movement of molecules will be continuous until the concentration of the same
molecule will be equal on either side of the membrane.

𝐶ℎ𝑖𝑔ℎ & 𝐶𝑙𝑜𝑤 are the concentration of the molecules on the side one and side two
respectively of a cell membrane. Here the free energy change on side 1 and side 2
is driven as:
[𝐶𝐻𝑖𝑔ℎ ]
∆G = −RT In
[𝐶𝑙𝑜𝑤 ]
[𝐶𝐻𝑖𝑔ℎ ]
∆G = −2.303 RT log
[𝐶𝑙𝑜𝑤 ]

The difference in the concentration (𝐶ℎ𝑖𝑔ℎ - 𝐶𝑙𝑜𝑤 ) is termed as the concentration

gradient and the ∆𝐺 is the chemical potential difference.

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Passive transport of charged molecules
The movement of charged ions depends upon two factors (1) Electric (Voltage)
Gradient-The difference in voltage across the membrane (2) Chemical
(concentration) Gradient. The difference in concentration across the membrane. In
combined form these two forces jointly named as Electrochemical gradient, that
determines the energetically favourable direction of transport of a molecule. The
movement of charged molecules is somewhat more difficult because the net charge
they carry affects the ability of a membrane to permit the transport.

Here Z is the charge on the transported species, F is the Faraday's constant (the
charge on 1 mole of electrons=96,485 coulombs/mol =96,485 joules/volt mol,
because 1 volt = 1 joule/coulomb) and AV is the electric potential difference (in
voltage difference) across the membrane.

In a case, if the sum of two terms on the right side of the equation is a negative
number, then the transport of the ion from side one to side two would occur
spontaneously.
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Passive transport always depends on the following factors:
1. Substrate concentration gradient across the layer.
2. Hydrophobicity of the molecule to be transported.
3. Size and charge of the molecule.
4. Electric potential across the membrane.
5. Diffusion area and distances.

Hydrophobicity of a substance is measured by its partition coefficient K. K is the

equilibrium for its partition between oil and water. The substance with high
partition coefficient is more soluble in lipid. For example, diethyl urea having an
ethyl group (CH3CH2O) attached to each nitrogen atom of urea, has a K of 0.01,
whereas urea has a K of 0.0002. Diethyl urea is 50 times (0.01/0.0002) more
hydrophobic than urea so it will diffuse approx 50 times faster than urea.

Facilitated Diffusion of uncharged molecule/polar molecule

Facilitated diffusion is a spontaneous process of passive diffusion of a molecule or
an ion across living membranes with the help of specific membrane integral
proteins, containing the multiple membranes spanning a helices. These proteins
have two common features:
(i) They facilitate net movement of solutes only in the thermodynamically
favourable situation (that is AG<0).
(ii) They show a measurable affinity and specificity for the transported solute.
These proteins also show the behaviour of saturation regarding their binding
with solute molecules that experimentally distinguish the facilitated diffusion
with simple diffusion.

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Many proteins facilitating the diffusion by forming a protein-lined pathway through
the membrane. They allow the movement of hydrophilic molecules without their
contact with a hydrophobic interior of the membrane.

Transporters
The transporter proteins are also known as the carriers, they move a wide variety
of ions or molecules. A carrier protein alternates between two conformations,
likewise there are three types of transporters have been identified.

The rate of facilitated diffusion by uniporters is very high than passive diffusion
through a pure phospholipids bilayer and there is a maximum transport rate Vmax
that is achieved when the concentration gradient across the membrane is very high
and each uniporter is working at a maximal rate. The transport is also specific
because each uniporter transports only a single type of molecule or a single group
of closely related molecules.

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GLUT 1-GLUT 12 are the 12 glucose transporters expressed in humans and the
structure of all GLUT isoforms are quite similar with 12 hydrophobic membrane-
spanning a-helices. Many helices having the amino acids (e.g. serine, threonine,
glutamine and asparagine) which can form hydrogen bonds with the OH group of
glucose. All GLUT proteins transport glucose but their differential expression among
the various body cells enables them to regulate glucose metabolism independently
at different rates.

For instance, GLUT 1 and GLUT3 are found in erythrocytes and other cells to take
up glucose continuously at high rates from the blood. GLUT 2 expressed in the liver
have very high influx efficiency in the response of insulin and here the excess
glucose is stored as a polymer, named glycogen. GLUT4 is expressed only in fat and
muscle cells which respond to insulin by increasing their glucose uptake.

The kinetics of glucose transport via GLUT1 uniporter is likely similar to an enzyme-
catalyzed chemical reaction. If the substrate glucose, S, is present initially only on
The outside of the membrane. In this case,

Here Sout-GLUT1 represents GLUT1 in the outward facing conformation of GLUT1

with bound glucose. As Michaelis Menten equation we can derive the following
expression for V0, the initial transport rate for Scatalyzed by GLUT1:

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Here S is the concentration of Solute (the initial concentration of Sin = 0). Vmax the
highest transport rate when all molecules of GLUT1 bound with the substrate,
occurs at an infinitely high substrate concentration outwards.

The value of Km of GLUT 1 for D-glucose is 1.5mm, where approximately half of the
total transporters have a glucose molecule bonded to the binding site and the
greater the transport rate Vmax will be half. Some isomers of glucose are also
transported by GLUT1 which have much lower Km like D-mannose (20mm) and D-
galactose (30mm). Thus GLUT1 is very specific with higher affinity for D-glucose.

Facilitated Diffusion of ions by channel proteins

Channels are hollow membrane-bound proteins having a hydrophilic gateway.
Channels are integral proteins embedded cell membrane. Ion channels control the
movement of ions through the cell membrane

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The channels are:
1. Selective for particular ions.
2. Involve in Facilitated diffusion of ions.
3. Regionally located at a specific site on a cell surface. Like Na channel at the soma
of neurons.
4. Functionally unique.

Channel selectivity depends on various factors:

1. The charge on the ion-that is, whether it is positive or negative.

2. On the size of the ion.

3. On how much water the ion attracts and holds around it.

Types of lon channels

 lon channels are either active or passive: Passive lon Channels are always open.
Passive channels, also called leakage channels, are always open and ions pass
through them continuously.
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