BIOLOGICAL MEMBRANES

Objectives

Importance Composition & Models Important properties Specialized structures RBC membrane Biomembranes Transport mechanisms

Importance/ Biological role

Maintenance of shape Control of movement of molecules across Cell-cell recognition and communication Cellular movement

Chemical composition

Lipids

Phospholipids: most predominant

Phosphatidylcholine (lecithin) : 40-50% Sphingomyelins Glycolipids: 2-10% absent in prokaryotes Most rigid lipid in membrane

Sphingolipids:

Cholesterol: free and esterified

Proteins

Inner mitochondrial membrane contains highest proportion of proteins

Carbohydrates do not exist in free form

Glycoproteins

Membrane proteins

Enzymatic activity:

Na+ - K+ ATPase pump

Translocases IP3 and DAG Fibronectin, Spectrin, Ankyrin Ion channels

Carrier proteins:

Signal transduction:

Interactions with ECM and cytoskeleton

Regulation of permeability

Membrane proteins
Peripheral proteins

Integral proteins

Bound to either face by electrostatic interactions/ hydrogen bonds

Released by mild treatment Salt solution of different ionic strength pH alteration

Usually enzymes Removal does not damage integrity

Embedded deeply (transmembrane) by hydrophobic bonds/ van der Waals force Removal requires use of detergents or organic solvents Transport proteins Removal causes denaturation of protein and loss of function

Lipid bilayer: Models

Amphipathic molecules Polar heads point outside, Non-polar tails point inwards Non-polar core acts as diffusion barrier: impermeable to polar molecules and ions 1925, Gorter and Grendel proposed lipid bilayer model 1935, Davson and Danielle suggested phospholipids as major constituent

Fluid mosaic model

1972, Singer and Nicholson Intrinsic proteins immersed in protein bilayer (60-100A) Extrinsic proteins loosely attached to surface of membrane Charecteristics:
Icebergs

in sea No flip flop

Fluid mosaic model

Membrane asymmetry

Proteins are inserted in asymmetric fashion Oligosaccharide units project towards exterior Lipids are distributed asymmetrically:
Outer

leaflet:

Phosphatidylcholine Sphingolipids

Inner

leaflet:

Phosphatidylethanolamine
Phosphatidylserine

Membrane fluidity

Temperature determine fluidity

Temp : fluidity Phase transition / melting temperature (Tm)

Composition
Short

chain FA fluidity Cis- Unsaturated FA fluidity (more the no. of double bonds, the Tm)

Cholesterol has dual role

decreases fluidity

above Tm Increases fluidity below Tm by acting like impurity

RBC membrane

Glycophorin
Integral

glycoprotein with oligosaccharide units facing exteriorly Determine antigen specificity

Anion exchanger
Integral

glycoprotein Extrusion of bicarbonate ions in exchange of chloride

Ankyrin
Peripheral

protein on cytoplasmic side Cross-linked to spectrin and actin

Other Biomembranes

Micelles: when critical concentration of lipids is present Liposomes

formed by sonication spheres of lipid bilayers that enclose aqueous medium.

Clinically
carriers

useful

of drugs in the circulation targeted therapy

Aquasomes

Most recently developed delivery system Used for proteins and peptides Structure: nanoparticulate, three layered, self assembled Central solid nanocrystalline core coated with polyhydroxy oligomers Biochemically active molecules adsorbed to this core Core gives structural stability and stabilizes active biological molecules Used for: Insulin, Hemoglobin, Antigens, Serratiopeptidase

Transport across cell membranes

Lipid bilayers are semipermeable

Non lipid-soluble molecules are handled by membrane proteins: Channels for ions/ small mols Transporters for larger mols

Lipid-protein association in membrane

Membrane phospholipids are solvents for membrane proteins - helical structures in proteins minimize hydrophilic character of peptide bonds Proteins are amphipathic Hydropathy plot

Helps to predict whether a protein can have transmembrane location

Specialized membrane structures

Lipid rafts
Exoplasmic

location Cholesterol, Sphingolipids and proteins

Caveolae
Caveolin-1

protein Flask shaped indentations in cytosolic side

Tight junctions
Prevent

diffusion of macromolecules Located below apical surfaces, between cells Proteins: occludin, claudins

Transport processes

Passive diffusion Carrier mediated transport

Facilitated diffusion Active transport

Primary active transport Secondary active transport

Exocytosis and endocytosis Transport through gap junctions

Passive diffusion

Gases: Highly permeable Water: Moderately permeable Ions and large polar molecules: Impermeable

Carrier-mediated transport

Mediated through integral proteins

Permeases/

Porters/ translocases

Proteins are specific (GLUT) Can be inhibited by structural analogs

1,5-anhydroglucitol

Uniport Symport/ Antiport: obligatory simultaneous cotransport of another mol

Facilitated diffusion

Along concentration gradient, no energy consumed Transport protein hastens the process Mechanism:
oscillation

between two conformations: ping-pong Process is reversible Kinetics follow Michelis - Menten rate law

Egs.
Glucose

transporters: GLUT Chloride transporters: Cl- / HCO3- antiport and cystic fibrosis transmembrane conductance

Rate kinetics

Active transport

Solute moves against concentration gradient Expenditure of energy Primary active transport:
Energy

obtained from ATP hydrolysis: Na-K ATPase

Secondary active transport:

Substrate

molecule moves coupled to another ion down its concentration gradient: Na- glucose symport No ATP hydrolysis, energy derived from electrochemical gradient of the primary ion

Primary active transport

Sodium Potassium ATPase

Secondary active transport

Sodium Glucose symport

Ion channels

Transmembrane proteins Selective to certain ions Allow ionic transport at high rates Regulated Types:
Voltage

gated Ligand gated Mechanically gated

Affected by drugs

Trans-membrane pore systems

Ionophores

Aquaporins

Small cyclic organic molecules Shuttles for ions Eg. Valinomycin and Gramicidin, used as antibiotics

Tetrameric transmembrane proteins Permit passage of water only Mutations cause nephrogenic diabetes insipidus

Endocytosis

Exocytosis

Segments of plasma membrane invaginate and enclose small vol of ECF Phagocytosis and pinocytosis Primary and secondary lysosomes LDL mol and receptor internalized

Release macromolecules Involved in membrane remodelling Ca++ triggers exocytosis