E N D
Biological membranes &Transport Na+-K+ PUMP M.Prasad Naidu MSc Medical Biochemistry, Ph.D.Research Scholar
MEMBRANES • Plasma membrane is an envelop surrounding the cell. • Separates & protects the cell from the external hostile environment. • Besides being a protective barrier, PM provides a connecting system b/n the cell & its environment.
Chemical composition • Membranes are composed of lipids, proteins & carbohydrates. • Among the lipids, amphipathic lipids ( containing both hydrophilic & hydrophobic groups) namely PL, GL and Cholesterol are found in the animal membranes.
Structure of membranes • Lipid bilayer model-- Davson&Danielle(35) • Fluid mosaic model-Singer&Nicolson(1972 • FM model is more recent and acceptable • Thickness---5-8nm • A membrane is essentially composed of a lipid bilayer. • The hydrophobic (non-polar) regions face each other at the core of the bilayer while the hydrophilic (polar) regions fact outward • Globular proteins are irregularly embedded in the lipid bilayer.
Membrane proteins • Membrane proteins are categorized into two groups. • 1. Extrinsic (peripheral)membrane proteins • Eg: Cyt.c of mitochondria • 2. Intrinsic (integral)membrane proteins. • Eg: hormone receptors, Cyt.P450 • The membrane is asymmetric due to the irregular distributions of proteins.
Transport across membranes • The biological membranes are relatively impermeable. • The membrane therefore forms a barrier for the free passage of compounds across it. • 3 distinct mechanisms have been identified for the transport of solutes (metabolites) through the membrane. • 1. Passive diffusion • 2. Facilitated diffusion • 3. Active Transport
Passive diffusion • A simple process which depends on the concentration gradient of a particular substance across the membrane. • Passage of water & gases through the membrane occurs by passive diffusion. • The process does not require energy.
Facilitated diffusion • This is somewhat comparable with PD, since the solute moves along the concentration gradient (higher to lower) and no E is needed. • FD occurs through the mediation of carrier or transport proteins. • Specific carrier proteins for the tpt of glu, gal, leu, phe etc have been isolated and characterized.
Factors affecting rate of diffusion • 1. Permeability of the cell membrane • 2. Temparature • 3. Conc.gr / electrical gr of the substance across the cell membrane • 4. Solubility of the substance • 5. Thickness of the cell membrane • 6. Size of the molecules • 7. Size of the ions. • 8. Charge of the ions
Active transport • AT occurs against a conc. gradient & this is dependent on the supply of metabolic E (ATP) • This is also a carrier mediated process like FD. • Movement of substances against the chemical or electrical or electrochemical gradient is called AT. • It is like swimming in the opp direction of water flow in a river( also called uphill transport) • The E required is liberated mostly by break down of high E compounds like ATP.
Active transport • AT occurs with the help of carrier proteins as in the case of FD. (mech .is diff) • Each CP can carry only one substance or more than one across the membrane. • Those CPs transporting only 1 substance are called uniports/uniport pumps. • Those CPs carrying more than one substance are called symports/antiports
Mechanism of active transport • When a substance to be transported across the cell membrane comes near the cell, it combines with the CP of the membrane and a Sub-Pro Complex is formed. • Now this SP complex moves towards the inner surface of the membrane. • Now the substance is released from the CPs • The same CP moves back to outer surface of the membrane to transport another molecule of the substance.
Substances transported by AT • Both ionic form and non-ionic forms • Ionic forms:- Na+, K+, Ca2+, H+, Cl- & I- • Non-ionic forms:- Glu, AAs, Urea
Types of AT • 1. Primary AT:- In this type of AT, the E is liberated directly from the break down of ATP. • By this method Na+, K+, Ca2+, H+, Cl- are transported across the membrane • 2. Secondary AT:- When Na+ is transported by a CP, another substance is also transported by the same protein simultaneously, either in the same direction or in the opp direction. • This type of tpt of a sbustance with Na+ by means of a CP is called the secondary AT. • i)CP tpts 2 diff molecules in the same direction (symport) • Ii)CP tpts 2 diff molecules in opp.direction (antiport)
Primary active transport of Na+ & K+ (Na+-K+ Pump) • Na+ & K+ are transported across the membrane by means of common mechanism called Na+-K+ Pump. • This tpts Na+ from inside the cell to outside and K+ from outside into the cell. • This pump is present in all parts of the body. • Cells have a high intracellular K+ conc and a low Na+ conc. • This is essentially needed for the survival of the cells. • High cellular K+ is needed for the optimal glycolysis & for protein biosynthesis. • Further Na+ & K+ gradients across the membranes are needed for the transmission of nerve impulse.
CP of Na+-K+ Pump • The CP involved in Na+-K+ Pump has got 6 sites • 3 receptor sites for Na+ . These are on the inner (towards the cytoplasm) surface of the protein molecule • 2 receptor sites for K+. These are on the outer (towards extra cellular fluid) surface of the protein molecule • 1 site for the enzyme ATPase (mol.wt250000) which is near the sites for Na. • ATPase consists of 2α & 2β subunits and represented as (αβ)2.
Mechanism of action of Na+-K+ Pump • 3 Na ions from the cell get attached to the receptor sites of Na+ on the inner surface of the CP. • 2 K ions outside the cell bind to the receptor sites of K+ located on the outer surface of the CP • The binding of Na+ & K+ to the CP immediately activates the ATPase. • ATPase causes the break down of ATP to ADP with the release of one high E phosphate.
Mechanism of action of Na+-K+ Pump • Now the E liberated causes some sort of conformational change in the CP. • Bcos of this, the outer surface of the CP with K+ , now faces the inner side of the cell. • And the inner surface of the CP with Na+ faces the outer side of the cell. • Now dissociation and release of the ions takes place so that the Na+ are released out side the cell and the K+ are released inside the cell. • The exact mechanisms involved in the dissociation and release of the ions are not yet known.
Electrogenic activity of Na+-K+ Pump • Na+-K+ Pump moves 3 Na+ outside the cell and 2 K+ inside the cell. • Thus, when the pump works once, there is a net loss of one +vely charged ion from the cell. • The continuous activity of these Na+-K+ Pump causes reduction in the no.of +vely charged ions inside the cell. • Leading to the development of –ve potential inside the cell. • This is called electrogenic activity of Na+-K+ Pump.