Cell Membrane

Dr.Urvashi Kumar Kohli

Cell Membrane
crucial to the life of the cell

encloses the cell,

defines its boundaries, and

maintains the essential differences between the cytosol and the extracellular environment.

1.1

Functions and properties of membrane

Membrane is a selectively permeable barrier between the cell and the external environment. Its function is to maintain Homeostasis. Its selective permeability allows the cell to maintain a constant internal environment. Plasma membranes form compartments (compartmentation) within cells.

Functions of Cell Membrane are to:

1. 2. 3.

4.

regulate cell volume , (control water in/ out) maintain intracellular pH (H+ regulation) selectively regulate ionic composition (e.g. Na+ , K+ ) concentrate metabolic fuel (nutrients, ATP, etc)

Membrane transport system ought to (continued):

5. concentrate and move building blocks (amino acids, etc) 6. remove toxic compounds (detoxification, trap and/or pump out) 7. generate ionic gradients to maintain excitability of nerve and muscle cells 8. control the flow of information within the cell, between cells and their environment.

A BRIEF HISTORY OF STUDIES ON PLASMA MEMBRANE STRUCTURE

Ernst Overton(1890s) Chemical nature of the outer boundary layer To test the permeability of the outer boundary layer, Overton placed plant root hairs into hundreds of different solutions containing a diverse array of solutes. He discovered that the more lipid soluble the solute, the more rapidly it would enter the root hair cells Remarks------dissolving power of the outer boundary layer of the cell matched that of a fatty oil

E. Gorter and F. Grendel(1925)

Ratio of the surface area of water covered by the extracted lipid to the surface area calculated for the red blood cells from which the lipid was extracted varied between 1.8 to 1 and 2.2 to 1 and concluded that the plasma membrane contained a bimolecular layer of lipids, that is, a lipid bilayer

-- the polar groups of each molecular layer (or leaflet) were directed outward toward the aqueous environment

In 1935, Hugh Davson and James Danielli proposed that the plasma
membrane was composed of a lipid bilayer that was lined on both its inner and outer surface by a layer of globular proteins

Davson and Danielli suggested that, in addition to the outer and inner protein layers, the lipid bilayer was also penetrated by protein-lined pores, which could provide conduits for polar solutes and ions to enter and exit the cell.

Fluid-mosaic model proposed in 1972 by S. Jonathan Singer and Garth

Nicolson
The fluid-mosaic model of membrane structure as initially proposed by Singer and Nicolson in 1972.

A current representation of the plasma membrane showing the same basic organization as that proposed by Singer and Nicolson

All biological membranes have a common general structure:

Lipid and protein molecules, held together mainly by noncovalent interactions.
Characteristics: dynamic, fluid structures, and most of their molecules are able to move about in the plane of the membrane lipid molecules - double layer about 6 nm thick - bilayer provides the basic structure of the membrane - serves as a relatively impermeable barrier to the passage of most water-soluble molecules. Protein molecules - transport specific molecules - catalyzing membrane associated reactions, such as ATP synthesis - structural links - connect the membrane to the cytoskeleton and/or to either the extracellular matrix or an adjacent cell, - receptors Carbohydrates cell membranes are asymmetrical structures

Figure - 1. Three views of a cell membrane. (A) An electron micrograph of a plasma membrane (of a human red blood cell) seen in cross-section. (B and C) Schematic drawings showing twodimensional and three-dimensional views of a cell membrane.

(A). The Lipid Bilayer 50% of the mass lipid molecules in cell membranes are amphipathic

The Lipid Bilayer Is a Two-dimensional Fluid

Three types of lipids i. ii. iii. phosphoglycerides, sphingolipids, cholesterol

most abundant phospholipids(phosphoglycerides)

Phospholipids
(Amphipathic)

Sphingolipids
(amphipathic)

Figure -2. The parts of a phospholipid molecule. Phosphatidylcholine, represented schematically (A), in formula (B), as a space-filling model (C), and as a symbol (D). The kink due to the cis-double bond is exaggerated in these drawings for emphasis

Figure 10-3. A lipid micelle and a lipid bilayer seen in cross-section. Lipid molecules form such structures spontaneously in water. The shape of the lipid molecule determines which of these structures is formed. Wedge-shaped lipid molecules (above) form micelles, whereas cylindershaped phospholipid molecules (below) form bilayers.

Figure - 4 . Phospholipid mobility. The types of movement possible for phospholipid molecules in a lipid bilayer.

The Fluidity of a Lipid Bilayer Depends on Its Composition

Phase Transition - Hydrocarbon chains (short and Double bond) - cholestrol Inhibit phase transition

Composition A. phospholipids ( 4 major) - phosphatidylcholine, sphingomyelin, phosphatidylserine, and phosphatidylethanolamine. - inositol phospholipids B. Cholestrol C. Glycolipids

Figure - 5. Influence of cis-double bonds in hydrocarbon chains. The double bonds make it more difficult to pack the chains together and therefore make the lipid bilayer more difficult to freeze.

Figure - 6. The structure of cholesterol. Cholesterol is represented by a formula in (A), by a schematic drawing in (B), and as a space-filling model in (C).

Figure - 7. Cholesterol in a lipid bilayer. Schematic drawing of a cholesterol molecule interacting with two phospholipid molecules in one leaflet of a lipid bilayer.

The Lipid Bilayer Is Asymmetrical

Figure - 8. Four major phospholipids in mammalian plasma membranes. Note that different head groups are represented by different symbols in this figure and the next. All of the lipid molecules shown are derived from glycerol except for sphingomyelin, which is derived from serine

The Lipid Bilayer Is Asymmetrical

Figure - 9. The asymmetrical distribution of phospholipids and glycolipids in the lipid bilayer of human red blood cells. The symbols used for the phospholipids are those introduced in Figure- 8. In addition, glycolipids are drawn with hexagonal polar head groups (blue). Cholesterol (not shown) is thought to be distributed about equally in both monolayers.

Plasma membrane, are synthesized in the endoplasmic reticulum (ER), and it is here that the phospholipid asymmetry is generated by the phospholipid translocators in the ER that move specific phospholipid molecules from one monolayer to the other.

Sphingolipids (amphipathic)

Glycolipids Are Found on the Surface of All Plasma Membranes

5% of the lipid molecules in the outer monolayer most complex of the glycolipids, the gangliosides - sialic acid - most abundant in the plasma membrane of nerve cells, where they constitute 5-10% of the total lipid mass,

Figure 10-12. Glycolipid molecules. Galactocerebroside (A) is called a neutral glycolipid because the sugar that forms its head group is uncharged. A ganglioside (B) always contains one or more negatively charged sialic acid residues (also called N-acetylneuraminic acid, or NANA), whose structure is shown in (C). Whereas in bacteria and plants almost all glycolipids are derived from glycerol, as are most phospholipids, in animal cells they are almost always produced from sphingosine, an amino alcohol derived from serine, as is the case for the phospholipid sphingomyelin (see Figure- . Gal = g

functions of glycolipids In the plasma membrane of epithelial cells, -glycolipids protect the membrane from the harsh conditions (such as low pH and degradative enzymes). Charged glycolipids, such as gangliosides, (electrical effects): their presence will alter the electrical field across the membrane and the concentrations of ions - especially Ca2+ - at its external surface. Glycolipids may also play a role in electrical insulation, since in the myelin membrane, which electrically insulates nerve cell axons, the noncytoplasmic half of the bilayer is filled with them. They are also thought to function in cell-recognition processes.

Nature and Importance of the Lipid Bilayer

Table -1 .Approximate Lipid Compositions of Different Cell Membranes

The Nature and Importance of the Lipid Bilayer

1. diversity in composition in different bacteria---physical state depends upon lipid composition

2.Thickness - 60 Angstron

3.Continuous unbroken structure

4.self resealing property 5.flexible-shape changes (during locomotion n cell division) 6.lipid bilayer- facilitate regulated fusion and budding of membranes 7.maintaining the proper internal composition of a cell, 8. in separating electric charges across the plasma membrane 9. Self Assemble ( PC Liposomes )

2. Proteins
THE STRUCTURE AND FUNCTIONS OF MEMBRANE PROTEINS
- Asymmetry of Proteins orientation in Membrane(membrane "sidedness") - membrane protein- has a defined orientation relative to the cytoplasm. -Therefore properties of one surface of a membrane are very different from those of other surface the other surface Three Classes of Proteins a. Integral membrane Proteins b. Peripheral Proteins c. GPI anchored Proteins

2. Proteins
Membrane Proteins Can Be Associated with the Lipid Bilayer in Various Ways

Integral Membrane Protein

Figure - Six ways in which membrane proteins associate with the lipid bilayer. Most transmembrane proteins are thought to extend across the bilayer as a single helix (1) or as multiple helices (2); some of these "single-pass" and "multipass" proteins have a covalently attachedfatty acid chain inserted in the cytoplasmic monolayer (1). Other membrane proteins are attached to the bilayer solely by a covalently attached lipid - either a fatty acid chain or prenyl group - in the cytoplasmic monolayer (3) or, less often, via an oligosaccharide, to a minor phospholipid, phosphatidylinositol, in the noncytoplasmic monolayer (4). Finally, many proteins are attached to the membrane only by noncovalent interactions with other membrane proteins (5) and (6). How the structure in (3) is formed is illustrated in next Figure

Figure 10-14. The covalent attachment of either of two types of lipid groups can help localize a water-soluble protein to a membrane after its synthesis in the cytosol. (A) A fatty acid chain (either myristic or palmitic acid) is attached via an amide linkage to an amino-terminal glycine. (B) A prenyl group (either farnesyl or a longer geranylgeranyl group - both related to cholesterol) is attached via a thioether linkage to a cysteine residue that is four residues from the carboxyl terminus. Following this prenylation, the terminal three amino acids are cleaved off and the new carboxyl terminus is methylated before insertion into the membrane. The structures of two lipid anchors are shown underneath: (C) a myristyl anchor (a 14-carbon saturated fatty acid chain), and (D) a farnesyl anchor (a 15-carbon unsaturated hydrocarbon chain).

In Most Transmembrane Proteins the Polypeptide Chain Is Thought to Cross the Lipid Bilayer in an alpha-helical Conformation

Figure 10-15. A segment of a transmembrane polypeptide chain crossing the lipid bilayer as an alpha helix. Only the - carbon backbone of the polypeptide chain is shown, with the hydrophobic amino acids in green and yellow.

Porins Are Pore-forming Transmembrane Proteins That Cross the Lipid Bilayer as a beta- Barrel

Figure . The three-dimensional structure of a porin trimer of Rhodobacter capsulatus determined by x-ray crystallography. (A) Each monomer consists of a 16-stranded antiparallel bbarrel that forms a transmembrane water-filled channel. (B) The monomers tightly associate to form trimers, which have three separate channels for the diffusion of small solutes through the bacterial outer membrane. A long loop of polypeptide chain (shown in red), which connects two bstrands, protrudes into the lumen of each channel, narrowing it to a cross-section of 0.6 x 1 nm.

Synthesis of TMM (Asymmetry of proteins)

Figure 10-17. A typical single-pass transmembrane protein. Note that the polypeptide chain traverses the lipid bilayer as a right-handed alpha- helix and that the oligosaccharide chains and disulfide bonds are all on the noncytosolic surface of the membrane. Disulfide bonds do not form between the sulfhydryl groups in the cytoplasmic domain of the protein because the reducing environment in the cytosol maintains these groups in their reduced (-SH) form.

location of proteins- reflects the function IMM(TM)- Functions on both sides- transport molecules across it. e.g. cell-surface receptors-TM proteins. Peripheral membrane proteinse.g. protiens involved in intracellular signaling (cytosolic half). TMM(IMM) Unique Orientation(Text) ---non-Cytoplasmic, membrane- spanning domain and cytoplasmic domain

Membrane Proteins Can Be Solubilized and Purified in Detergents

Figure 10-18. A detergent micelle in water, shown in cross-section. Because they have both polar and nonpolar ends, detergent molecules are amphipathic.

Figure 10-19. Solubilizing membrane proteins with a mild detergent. The detergent disrupts the lipid bilayer and brings the proteins into solution as protein-lipid-detergent complexes. The phospholipids in the membrane are also solubilized by the detergent.

Figure 10-20. The structures of two commonly used detergents. Sodium dodecyl sulfate (SDS) is an anionic detergent, and Triton X-100 is a nonionic detergent. The hydrophobic portion of each detergent is shown in green, and the hydrophilic portion is shown in blue. Note that the bracketed portion of Triton X-100 is repeated about eight times

Figure . The use of mild detergents for solubilizing, purifying, and reconstituting functional membrane protein systems. In this example functional Na+-K+ ATPase molecules are purified and incorporated into phospholipid vesicles. The Na+-K+ ATPase is an ion pump that is present in the plasma membrane of most animal cells; it uses the energy of ATP hydrolysis to pump Na+ out of the cell and K+ in,

Cells Can Confine Proteins and Lipids to Specific Domains Within a Membrane

Figure . Diagram of an epithelial cell showing how a plasma membrane protein is restricted to a particular domain of the membrane. Protein A (in the apical membrane) and protein B (in the basal and lateral membranes) can diffuse laterally in their own domains but are prevented from entering the other domain, at least partly by the specialized cell junction called a tight junction. Lipid molecules in the outer (noncytoplasmic) monolayer of the plasma membrane are likewise unable to diffuse between the two domains; lipids in the inner (cytoplasmic) monolayer, however, are able to do so (not shown).

 Six major functions of membrane proteins: Transport Enzymatic activity Signal transduction Cell-cell recognition Intercellular joining Attachment to the cytoskeleton and extracellular matrix (ECM)

LE 7-9a

Enzymes

Signal

ATP

Receptor

Transport

Enzymatic activity

Signal transduction

LE 7-9b

Glycoprotein

Cell-cell recognition

Intercellular joining

Attachment to the cytoskeleton and extracellular matrix (ECM)

The Cell Surface Is Coated with Sugar Residues

Figure . Simplified diagram of the cell coat (glycocalyx). The cell coat is made up of the oligosaccharide side chains of glycolipids and integral membrane glycoproteins and the polysaccharide chains on integral membrane proteoglycans. In addition, adsorbed glycoproteins and adsorbed proteoglycans (not shown) contribute to the glycocalyx in many cells. Note that all of the carbohydrate is on the noncytoplasmic surface of the membrane

The Role of Membrane Carbohydrates in Cell-Cell Recognition

Cells recognize each other by binding to surface molecules, often carbohydrates, on the plasma membrane Membrane carbohydrates may be covalently bonded to lipids (forming glycolipids) or more commonly to proteins (forming glycoproteins) Carbohydrates on the external side of the plasma membrane vary among species, individuals, and even cell types in an individual

Plasma membrane is a semi-permeable (selective) membrane/ Membrane structure results in selective permeability
Small hydrophobic molecules: O2, CO2, N2, benzene Small uncharged polar molecules: H2O, ethanol, glycerol

Lipid bilayer

Larger uncharged polar molecules: glucose, amino acid, nucleotides

Ions: H+, Na+, HCO3- , K+, Ca+, Mg2+, CL- , etc.

Transporters or channels

The process of diffusion of water is called osmosis. But why some can diffuse while some cannot? What criteria are controlling the diffusion of molecules across the membrane ?

Plasma membrane is a semi-permeable (selective) membrane

The process of diffusion of water is called osmosis. But why some can diffuse while some cannot? What criteria are controlling the diffusion of molecules across the membrane ?

Protein - free Lipid Bilayers Are Highly Impermeable to Ions

Figure 11-1. The relative permeability of a synthetic lipid bilayer to different classes of molecules. The smaller the molecule and, more important, the fewer hydrogen bonds it makes with water, the more rapidly the molecule diffuses across the bilayer.

Simple diffusion : cell membranes allow water and non-polar molecules to permeate
Channel protein- Passive diffusion

Carrier Protein- Active or Passive

Two classes of membrane transport proteins

Figure 11-3. A schematic view of the two classes of membrane transport proteins. A carrier protein is thought to alternate between two conformations, so that the solute binding site is sequentially accessible on one side of the bilayer and then on the other. In contrast, a channel protein is thought to form a water-filled pore across the bilayer through which specific ions can Diffuse.

Figure 11-4. Comparison of passive transport down an electrochemical gradient with active transport against an electrochemical gradient. Whereas simple diffusion and passive transport by membrane transport proteins (facilitated diffusion) occur spontaneously, active transport requires an input of metabolic energy. Only carrier proteins can carry out active transport, but both carrier proteins and channel proteins can mediate facilitated diffusion.

Three types of carrier-mediated transport26

UniportGlucose carriers in animal cells take up glucose from extracellular fluid. (passive transport)

Symport -glucose with Na+ intestinal cells and Kidney cells take up glucose from the lumen of the intestine and kidney tubules(low)

Antiport Band 3 protein (anion carrier) In human RBC- exchange Clfor HCO3-)

Figure - Three types of carrier-mediated transport. The schematic diagram shows carrier proteins functioning as uniports, symports, and antiports.

Conformational change in a carrier protein could mediate the facilitated diffusion of a solute

Figure . A hypothetical model showing how a conformational change in a carrier protein could mediate the facilitated diffusion of a solute. The carrier protein shown can exist in two conformational states: in state "pong" the binding sites for solute A are exposed on the outside of the bilayer; in state "ping" the same sites are exposed on the other side of the bilayer. The transition between the two states is proposed to occur randomly and to be completely reversible. Therefore, if the concentration of A is higher on the outside of the bilayer, more A will bind to the carrier protein in the pong conformation than in the ping conformation, and there will be a net transport of A down its electrochemical gradient.

Table 11-1. Comparison of Ion Concentrations Inside and Outside a Typical Mammalian Cell

Pumping cycle of the Na+-K+ ATPase Primary Acitve Transport

Figure . A schematic model of the pumping cycle of the Na+-K+ ATPase. The binding of Na+ (1) and the subsequent phosphorylation by ATP of the cytoplasmic face of the ATPase (2) induce the protein to undergo a conformational change that transfers the Na+ across the membrane and releases it on the outside (3). Then the binding of K+on the extracellular surface (4) and the subsequent dephosphorylation (5) return the protein to its original conformation, which transfers the K+ across the membrane and releases it into the cytosol (6). These changes in conformation are analogous to the ping pong transitions shown in Figure - except that here the Na+- dependent phosphorylation and the K+-dependent dephosphorylation of the protein cause the conforma-tional transitions to occur in an orderly manner, enabling the protein to do useful work. Although for simplicity only one Na+- and one K+-binding site are shown, in the real pump there are thought to be three Na+- and two K+binding sites. Moreover, although the ATPase is shown as alternating between two conformational states, there is evidence that it goes through a more complex series of conformational changes during the actual pumping cycle.

Figure 11-10. The Na+-K+ ATPase. This carrier protein actively pumps Na+ out of and K+ into a cell against their electrochemical gradients. For every molecule of ATP hydrolyzed inside the cell, three Na+ are pumped out and two K+ are pumped in. The specific pump inhibitor ouabain and K+ compete for the same site on the external side of the ATPase.

Secondary Active Transport: Ion Gradients

Na+ Glucose HClO3Cl-

Figure 11-13. The transcellular transport of glucose across an intestinal epithelial cell

Cells maintain voltage across plasma membranes

Cells maintain voltage across plasma membranes. Cytoplasm negative compared to opposite side of membrane (membrane potential - ranges from -50 to -200 millivolts)

 Membrane potential favors passive transport of cations (positive ions) into cell and anions (negative ions) out of cell. Creates an electrochemical gradient across membrane.

 Some organisms have proton pumps that actively pump H+ out of cell (i.e. plants, bacteria, and fungi)

 Membrane potential is the voltage difference across a membrane Two combined forces, collectively called the electrochemical gradient, drive the diffusion of ions across a membrane: A chemical force (the ions concentration gradient) An electrical force (the effect of the membrane potential on the ions movement)

Maintenance of Membrane Potential by Ion Pumps

 An electrogenic pump is a transport protein that generates the voltage across a membrane The main electrogenic pump of plants, fungi, and bacteria is a proton pump

Ionophores

Gramicidin A

Valinomycin (K+)

Figure . A mobile ion carrier and a channel-forming ionophore. In both cases net ion flow occurs only down an electrochemical gradient.

Figure 11-6. The structure of a gramicidin channel. The channel is formed by the association of two identical peptides at their amino-terminal ends. Each chain is folded into a b helix, which resembles a rolled-up b pleated sheet. (A) is a side view and (B) a top view. The peptide backbones that line the channel are shown in blue and dark green, while the light green represents the protruding hydrophobic side chains. The lipid bilayer is shown in gray. (C) shows the size of unhydrated K+ions, while (D) shows a membrane-spanning a helix in top view for comparison with the b helix.

Bulk transport across the plasma membrane occurs by exocytosis and endocytosis
Small molecules and water enter or leave the cell through the lipid bilayer or by transport proteins Large molecules, such as polysaccharides and proteins, cross the membrane via vesicles

Exocytosis
In exocytosis, transport vesicles migrate to the membrane, fuse with it, and release their contents Many secretory cells use exocytosis to export their products

Endocytosis
In endocytosis, the cell takes in macromolecules by forming vesicles from the plasma membrane Endocytosis is a reversal of exocytosis, involving different proteins

 Three types of endocytosis: Phagocytosis (cellular eating): Cell engulfs particle in a vacuole Pinocytosis (cellular drinking): Cell creates vesicle around fluid Receptor-mediated endocytosis: Binding of ligands to receptors triggers vesicle formation

LE 7-20c

RECEPTOR-MEDIATED ENDOCYTOSIS
Coat protein Receptor Coated vesicle

Coated pit

Ligand
Coat protein A coated pit and a coated vesicle formed during receptormediated endocytosis (TEMs). 0.25 m

Plasma membrane

Synthesis and Sidedness of Membranes

Asymmetry of plasma membranes

1. 2. 3. 4. 5. Membrane separates interior and exterior side of the cell and the 2 faces of lipid bilayer are different in composition and structure, with different proteins and phospholipids. The plasma membrane core contain 1/3 cholesterol and 2/3 phospholipids and sphingolipids, the outer leaflet contains 5% glycolipids. Oligosaccharide chains are attached at outer face of lipids or proteins. Shingomyelin and phosphatidylcholine are mainly at the outer face of the bilayer. Phosphotidylethanolamine and phosphatidylserine are mainly in the inner face. Asymmetry of proteins- Oligosaccharides and Disulphide bonds in the protein on non-cytoplasmis side

6.

Chemical and physical properties of plasma membranes

1. Sheet-like structure consists of proteins, lipids, with carbohydrates attached to them. 2. Non-covalent assemblies of lipids and proteins. 3. The proteins serve the functions as transporters, channels, enzymes, signal transducers, etc. 4. The lipid molecules are amphipathic molecules, they have both hydrophilic (polar) and hydrophobic (non-polar) moieties. 5. Barriers to the flow of charged molecules.