Cell Membrane

Objective
To identify the nature of proteins and lipids that form the structural elements of biological membranes and their role in the biological activity of membranes. To identify the general features of the fluid mosaic model of membrane structure. To identify the classes of lipids found in biological membranes, their key structural features, and the most prominent members of each class. To identify the process of lipid self-assembly and discriminate between micelles and liposomes. To identify how hydrocarbon chain length, degree of unsaturation, and temperature impact the fluidity of a membrane. To identify the range of protein content of biological membranes and the role of proteins in the membranes. To identify the orientation of domains and the way that integral membrane proteins interact with other proteins and with the lipid bilayer. To identify the basis for the constant asymmetry found in the distribution of lipids and proteins of the inner and outer monolayers of a given biological membrane. To identify the difference between simple diffusion and facilitated diffusion. To identify the difference between facilitated diffusion and active transport. To identify the similarities and differences between symports and antiports.

Membranes serve a number of essential cellular functions. They constitute the boundaries of cells and intracellular organelles, and they provide a surface where many important biological reactions and processes occur. Membranes have proteins that mediate and regulate the transport of metabolites, macromolecules, and ions. Hormones and many other biological signal molecules and regulatory agents exert their effects via interactions with membranes. Photosynthesis, electron transport, oxidative phosphorylation, muscle contraction, and electrical activity all depend on membranes and membrane proteins. Thirty percent of the genes of at least one organism, Mycoplasma genitalium (whose entire genome has been sequenced), are thought to encode membrane proteins. Biological membranes are uniquely organized arrays of lipids and proteins (either of which may be decorated with carbohydrate groups). The lipids found in biological systems are often amphipathic, signifying that they possess both polar and nonpolar groups. The hydrophobic nature of lipid molecules allows membranes to act as effective barriers to polar molecules. The polar moieties of amphipathic lipids typically lie at the surface of membranes, where they interact with water. Proteins interact with the lipids of membranes in a variety of ways. Some proteins associate with membranes via electrostatic interactions with polar groups on the membrane surface, whereas other proteins are embedded to various extents in the hydrophobic core of the membrane. Other proteins are anchored to membranes via covalently bound lipid molecules that associate strongly with the hydrophobic membrane core.

Importance of Cell Membrane

Limits or protects cell by maintaining homeostasis by acting as a selective barrier so as to affect the cell s composition, concentration, osmotic pressure or volume. Model for other intracellular structures. Plays a role in transport mechanism. Plays a role in cell to cell recognition and also serves as receptors for hormones, drugs and viruses. Can be used to differentiate normal cells from cancerous cells in microscopic examination.

General Features of Cell Membrane

Cell membrane is formed by a continuous bilayer of lipids which protein complexes are embedded in it. There are other proteins peripheral to the lipid bilayer disposed on the inner surface. The membrane is highly assymetrical. The asymmetry is further emphasized in that the chains oligosaccharide protrude only at the outer surface of the membrane.

Composition of Cell Membrane

Protein: 52% Lipids: 40% CHO: 8% (glycolipid and glycoprotein)

LIPIDS

Lipids
in cell membrane shows the property of sefassembly, self-resealing and fluidity. The fluidity in turn is affected by temperature, composition of hydrocarbon chain, and the cholesterol content.

Composition of Lipid Fraction (Human RBC)

Membrane Lipid 1. Phosphoglycerides a. Phoshatidyl choline (17%) b. Phosphatidyl ethanolamine 16% c. Phosphatidyl serine (6%) 2. Sphingomyelin (16%) Hydrophobic Unit Fatty acid chain Hydrophilic Unit Phosphorylated alcohol

Fatty acid chain, Hydrocarbon chain of sphingosine Entire molecule except OH group Fatty acid chain, Hydrocarbon chain of sphingosine

Phosphoryl choline

3. Cholesterol (23%) 4. Glycolipid

OH group at Carbon 3 Sugar residues

Characteristics Features of Lipid in Human RBC

1. Lipids that terminate in CHOLINE are in the outer half while most of the phospholipids that contain terminal amino groups are in the inner half of the lipid bilayer. Therefore, phosphatidyl choline and sphingomyelin are more abundant outside while phosphatidyl ethanolamine and phosphatidyl serine are more abundant inside. Lipid molecules cannot flip flop between monolayers. Glycolipids constitute 5% of the outer monolayer and they are derived from sphingosine and are called GLYCOSPHINGOLIPID. Lipids in water will form spherical molecules called micelles or will form lipid bilayers, hiding the tail (non-plar) from the aqueous media. PHASE Transition: change from liquid phase to solid- hexagonal lattice network (gel-phase) Cholesterol are found outside the RBC. The OH group is attached to the polar head, and the steroid nucleus is attached to the hydrocarbon chain. It adds flexibility to the chain, and in so doing, prevent gelling.

2. 3.

PROTEINS

Proteins
Plays a role in mechanical structure Serves as carrier or channel for transport Involved in regulatory or ligand recognition property

Membrane protein can be classified into two categories

Extrinsic or peripheral proteins: proteins that are loosely bound to the membrane surface and can be easily removed. Intrinsic or integral proteins: make up 70% or more of total membrane protein. They are tightly bound to the lipid portion and can be removed only by drastic treatment.

Extrinsic Protein 1. Requirement for dissociation from membrane 2. Association with lipid when solubilized 3. Solubility after dissociation from membrane 4. Example Mild treatment; high ionic strength; metal ion chelating agents Free of lipid Soluble and molecularly dispersed in neutral aqueous buffer Spectrin, Antyrin

Intrinsic Protein Hydrophobic bond breaking agents; organic solvents; detergents Usually associated with lipid Usually insoluble and aggregated in neutral aq. buffer Polypeptide 3 Glycophorin Most membrane bound enzymes; drugs and hormonal receptors HLA

PROTEINS IN THE RBC

Peripheral Proteins
Polypeptide 1 and 2 (spectrin, tektin A) Protein Bond 5 (actin) Protein Bond 2.1, 2.2, 2.3, and 2.6 (ankyrin) Protein 4.1, 4.2, 6 and 7

Integral Proteins
Ion exchange protein or Bond 3 or component A Glycophorin

Note
Spectrin, actin and protein 4.1 forms the supra-molecular filaments of cell membrane. It maintains its shape and strength. Ankyrin anchors spectrin to the basement membrane. It is also attached to Bond 3 in the basement membrane. Bond 3: the COOH terminal ends at the outer aqueous layer while the NH2 terminal ends in cytoplasmic surface for binding of larger proteins such as ankyrin. It is an anionic binding polypeptide with binding sites for sialic acid. Glycophorin: differs from Bond 3 in that it is the NH2 that is exposed to the outer aqueous layer. It is responsible for cell to cell recognition, binding of receptors for blood group antigens, bacteria and viruses.

Membrane Molecular Models

Danieli Davson (1935) proposed that a lipid bilayer had protein adhering to both lipid aqueous surfaces. Robertson (1960) led the concept of a trilayered plasma membrane known as UNIT MEMBRANE. Based on electron microscopy, the dense outer layer corresponds to protein and the less dense middle layer corresponds to the hydrocarbon of the lipid. Benson (late 1960) suggested that protein layer existed as globular subunits within the biomolecular lipid layer with cost of the protein in alpha-helical conformation rather than continued layer inbeta conformation.