Bhaskar Ganguly Ph.D. Scholar (Vety. Biochemistry) Animal Biotechnology Laboratory C.V.A.Sc., G.B.P.U.A. & T.

Pantnagar, INDIA

 Overton {1890s}:
Lipids are present on cell surfaces; cell coats are probably mixtures of cholesterol & lecithin

Langmuir {1900s}:
Phospholipid monolayer

Gorter & Grendel {1925}:

Lipid Bilayer

 Davson & Danielli {1935}:

Biological membranes consist of lipid bilayers coated on both sides by thin sheets of proteins

Robertson {1960}:
All cellular membranes share a common underlying structure viz. Unit Membrane

 Singer & Nicolson {1972}:

Biological membranes consist of a mosaic of proteins in a lipid bilayer; The Fluid-Mosaic Model

 Behavior of lipid matrix depends on the properties of individual lipid components

Lipid matrix interacts with proteins and influences activity of the proteins Depending on duration of interactions, lipids are classified as Restricted lipids: long residence time, slow exchange with surrounding lipids Interfacial lipids: form coat or ring around the circumference of proteins, exchange rapidly with surrounding lipids

Restricted and Interfacial lipids may be necessary for protein function

 Form structural and environmental framework for cell function Phosphatidylcholine (PC), phosphatidylserine (PS) & phosphatidylinositol (PI): provide hydrated or charged membrane surfaces, allowing water/ ions to bind Phosphatidylethanolamine (PE): hydrophobic, promotes surface interactions without protein-protein interactions, promotes formation of non-bilayer structures; necessary for membrane fusion
Asymmetrical distribution between inner and outer leaflets Outer leaflet: rich in PC & sphingomyelin Inner leaflet: rich in PE & PS

Asymmetrical distribution is achieved & maintained by ATPdependent Aminophospholipid Translocase; translocates PE & PS between leaflets

Differential distribution of lipids in leaflets

Per cent distribution of phospholipids in erythrocyte membrane

Role of translocases/ flippases in maintaining membrane asymmetry

 Reduces freedom of movement of phospholipids, rigidifying effect on membrane viz. Condensing effect Non-uniform distribution in different cell membranes Decreases fluidity at high temperatures; increases fluidity at low temperatures
Decreases permeability to ions & small polar molecules

At low temperatures, cholesterol disallows close packing of hydrocarbon chains; at high temperatures, the rigid molecule restricts freedom of the acyl chains

 A state of change achieved by the motions of individual membrane components & their interrelationships in nonrepeating units of the membrane

Asymmetric distribution of different lipids adds another dimension to Membrane Dynamics

Re-distribution (lateral &/ or transverse) influences membrane properties, and allows differential regulation of membrane proteins Biological case studies:

PLATELETS, & PHOTORECEPTORS

 PC & Sphingomyelin in outer leaflet; PE & PS in inner leaflet Cholesterol : Phospholipid 0.50
Platelets cannot synthesize cholesterol; derived from megakaryocyte progenitor

Membrane cholesterol concentration represents plasma cholesterol concentration

Membrane cholesterol is also acquired from plasma lipoproteins

 Upon stimulus for aggregation, asymmetry of phospholipid distribution is lost

PE is rapidly translocated from inner to outer leaflet Aminophospholipid translocase is not inhibited; instead, Scramblase is involved (induced by high intracellular Ca++) Cholesterol translocates from outer to inner leaflet; thermodynamic exclusion of cholesterol due to unfavorable entropy of co-existence with PE Higher cholesterol results in stronger response to stimulus High membrane cholesterol platelets are more sensitive to epinephrine, ADP, collagen & thromboxane A2 Cholesterol enrichment increases signaling events viz. release of arachidonic acid, increased adrenergic & thrombin receptors, and higher Ca++ & inositol phosphate levels

 Cholesterol behaves both as a restricted and interfacial lipid Platelet stimulation increases rigidity & decrease fluidity
Activation of platelets alters platelet membrane to create a catalytic site for conversion of factor X to factor X-a, and of prothrombin to thrombin, leading to fibrinogen formation Creation of catalytic site requires surfacing of PS from the inner leaflet

Rod Outer Segment (ROS)

Plasma Membrane

Disk

Biochemical events initiating the impulse occur in membranous sacs called disks in the ROS New disks form from the ROS plasma membrane; old disks displaced apically are shed off & phagocytosed by retinal epithelium ( 10 days)

 Disks & Plasma membranes differ in lipid composition

Plasma membrane: richer in cholesterol & squalene (sterol precursor), PE:PC = 0.16, docosahexaenoic acid (DHA) ~5 % Disks: PE:PC = 0.92, DHA ~ 35 %

11-cis retinal all trans retinal

Rhodopsin Metarhodopsin I Metarhodopsin II

Metarhodopsin II Transducin cGMP dependent Phosphodiesterase (PDEase)

Basal disk ~ 30 mol% cholesterol, apical disk ~ 5 mol%; same mechanism as platelets (exclusion from PE-rich disk membrane) Cholesterol influences rhodopsin function; cholesterol inhibits activation of PDEase by rhodopsin
Conversion of Metarhodopsin I to larger Metarhodopsin II requires kinking of unsaturated acyl chains; cholesterol resists these free volume changes

 Cholesterol also interacts directly with rhodopsin DHA influences regeneration of rhodopsin

DHA can contribute as six cis- bonds; cis- bonds increase kinking within membrane bilayer thereby increasing free volume Rhodopsin is maintained in a relatively inactive state in plasma membrane; cholesterol (= inhibition of activation), DHA (= slow regeneration), i.e., Cholesterol/DHA favors inactivity In disk membrane, Cholesterol/DHA favors rapid activation and regeneration necessary for proper vision

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