Department of Natural Sciences University of St.

La Salle Bacolod City

CELL MEMBRANE or PLASMALEMMA

 Plasma membrane is present in both eukaryotic and prokaryotic cells.  Despite their differing functions, all biological membranes have a common general structure: each is a very thin film of lipid and protein molecules, held together mainly by noncovalent interactions.  Too thin to be seen with the light microscope, but in EM, the membrane is 8.5-10 nm thick and shows the trilaminar structure of the unit membrane.  Specialized techniques, such as x-ray diffraction and freeze-fracture electron microscopy, are needed to reveal the details of its organization.

STUDYING CELL MEMBRANES

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

Freeze-fracture EM provides images of both the hydrophobic interior of the cytosolic half of the bilayer (the P face) and the hydrophobic interior of the external half of the bilayer (the E face). After the fracturing process, the exposed fracture faces are shadowed with platinum and carbon, the organic material is digested away, and the resulting platinum replica is examined in EM.

Models of Plasma Membrane Structure

 Danielli and Davson model 2 layers of protein on either side of a bimolecular leaflet of mixed lipid.  Fluid-mosaic model- phospholipid bilayer with proteins floating on a sea of phospholipids.

 There are 3 major classes of membrane lipid molecules phospholipids, cholesterol, and glycolipids. The most abundant are the phospholipids.  Lipid molecules in cell membranes are amphipathic or amphiphilic i.e., they have a hydrophilic or polar end and a hydrophobic or nonpolar end.

 Hydrophilic molecules dissolve readily in water because they contain charged or uncharged polar groups that can form either favorable electrostatic interactions or hydrogen bonds with water molecules.  Hdrophobic molecules are insoluble in water because almost all of their atoms are uncharged and nonpolar and therefore cannot form energetically favorable interactions with water molecules.  If dispersed in water, they force the adjacent water molecules to reorganize into icelike cages that surround the hydrophobic molecule.  Because these cage structures are more ordered than the surrounding water, their formation increases free energy.  This free energy cost is minimized if the hydrophobic portions of amphipathic molecules cluster together so that the smallest number of water molecules is affected.

How hydrophilic and hydrophobic molecules interact differently with water. (A) Because acetone is polar, it can form favorable electrostatic interactions with water molecules, which are also polar. Thus, acetone readily dissolves in water. (B) Insoluble molecules such as 2-methyl propane is entirely hydrophobic & cannot form favorable interactions with water, forcing adjacent water molecules to reorganize into icelike cage structures, which increases the free energy. The symbol - indicates a partial negative charge, and + indicates a partial positive charge. Polar atoms are shown in color and nonpolar groups are shown in gray.

 Lipid composition can influence the activity of particular membrane proteins. They can determine the physical state of the membrane.  Depending on their shape, lipid molecules can form spherical micelles, with the tails inward; or they can form bimolecular sheets, or bilayers, with the hydrophobic tails sandwiched between the hydrophilic head groups.  Being cylindrical, phospholipid molecules spontaneously form bilayers in aqueous environments.  A lipid bilayer is responsible for the fluid characteristic of the membrane.  In this energetically most-favorable arrangement, the hydrophilic heads face the water at each surface of the bilayer, and the hydrophobic tails are shielded from the water in the interior.

(A) Wedge-shaped lipid molecules form micelles, whereas cylinder-shaped phospholipid molecules form bilayers. (B) A lipid micelle and a lipid bilayer seen in cross section.

PHOSPHOLIPIDS: have a polar head group and 2 hydrophobic hydrocarbon tails. The tails are usually fatty acids, differing in length from 14-24 C. Unsaturated FA have cis-double bands, producing a kink in the tail. Differences in length and saturation influence the packing of phospholipids , affecting the fluidity of the membrane.

Types of phospholipids: phosphoglycerides have a glycerol backbone; sphingolipids contain sphingosine instead of glycerol.

 The same forces that drive phospholipids to form bilayers also provide a self-healing property.  A small tear in the bilayer creates a free edge with water; because this is energetically unfavorable, the lipids spontaneously rearrange to eliminate the free edge. (In eukaryotic plasma membranes, larger tears are repaired by the fusion of intracellular vesicles.)

The only way for a bilayer to avoid having free edges is by closing in on itself and form a sealed compartment. The closed structure is stable because it avoids the exposure of the hydrophobic hydrocarbon tails to water, which would be energetically unfavorable.

Phospholipids very rarely migrate from the monolayer on one side to that on the other ("flip-flop). In contrast, lipid molecules readily exchange places with their neighbors within a monolayer giving rise to a rapid lateral diffusion.

Cholesterol reduces membrane fluidity at moderate temperatures by reducing phospholipid movement, but at low temperatures it hinders solidification by disrupting the regular packing of phospholipids.

Eukaryotic plasma membranes contain up to one molecule of for every phospholipid molecule which enhance the permeability-barrier properties of the lipid bilayer.

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

 They orient themselves in the bilayer with their hydroxyl groups close to the polar head groups of the phospholipid molecules. In this position, their rigid, platelike steroid rings interact with and partly immobilize those regions of the hydrocarbon chains closest to the polar head groups. By decreasing the mobility of the first few CH2 groups of the hydrocarbon chains of the phospholipid molecules, cholesterol makes the lipid bilayer less deformable in this region and thereby decreases the permeability of the bilayer to small water-soluble molecules. Although cholesterol tends to make lipid bilayers less fluid, at the high concentrations found in most eukaryotic plasma membranes, it also prevents the hydrocarbon chains from coming together and crystallizing. In this way, it inhibits possible phase transitions.

Spur cell anemia is caused by too much cholesterol in RBC membrane. It is common in alcoholics with cirrhosis of liver. Serum cholesterol gets very high and goes into the outer leaflet of RBC membrane. This expands the outer leaflet and produce spurs. The membrane loses flexibility and cells become round and spur covered. The cells get trapped in spleen capillaries and are destroyed. Total RBC count decreases resulting to anemia.

GLYCOLIPIDS: (A) Galactocerebroside, a neutral glycolipid because the sugar that forms its head group is uncharged. (B) A ganglioside always contains one or more negatively charged sialic acid residues (also called N-acetylneuraminic acid, or NANA) shown in (C). Almost all glycolipids are derived from glycerol, as are most phospholipids; in animal cells they are produced from serine, e.g. phospholipid sphingomyelin. 3 uncharged sugars: Gal = galactose; Glc = glucose, GalNAc = N- acetylgalactosamine

 Glycolipids are confined to the exposed apical surface, where they help protect the membrane against the harsh conditions such as low pH and degradative enzymes. Charged glycolipids, such as gangliosides, alter the electrical field across the membrane and the concentrations of ions especially Ca+2 at the membrane surface. They function in cell-recognition processes, in which membrane-bound carbohydrate-binding proteins (lectins) bind to the sugar groups on both glycolipids and glycoproteins in the process of cell-cell adhesion Mutant mice that are deficient in all of their complex gangliosides show no obvious abnormalities, although the males cannot transport testosterone normally in the testes and are consequently sterile.

Tay-Sachs disease is a fatal inherited condition that occurs when harmful quantities of gangliosides accumulate in the nerve cells of the brain because of deficiency in -Nacetylhexosaminidase A, an enzyme which catalyzes the One of the symptoms of Taybiodegradation of Sachs disease is the reddish gangliosides, eventually dot in the retina of the eye leading to the premature death which was first noticed by Warren Tay in the year 1881. of those cells. The disease is caused by the genetic mutation of HEXA gene on chromosome 15 which provides instructions for making the GM2 activator protein, a cofactor required for normal function of the -N-acetylhexosaminidase A.

Different mixtures of lipids are found in the membranes of cells of different types, as well as in the various membranes of a single eucaryotic cell. Functions include: 1. Certain membrane proteins accumulate in lipid rafts which are specialized areas in membranes where some lipids (primarily sphingolipids and cholesterol) and proteins (green) are concentrated. Rafts concentrate them for transport in small vesicles or to enable the proteins to function together, such as when they convert extracellular signals into intracellular ones.

Caveolae- special type of lipid rafts specialized for endocytosis and compartmentalization of signal transduction at the cell surface. One pathway is through formation of vesicles that bud off from the PM and travel to endosomes via microtubules. Another pathway is through cyclic budding at the cell surface to deliver small molecules and ions. The 3rd type form tubules from the cell surface to the center.

2. Some membrane-bound enzymes require specific lipid head groups in order to function.

3. The head groups of some lipids form docking sites for specific cytosolic proteins.

4.Some extracellular signals that act through membrane receptor proteins activate phospholipases that cleave selected phospholipid molecules in the plasma membrane, thereby generating fragments that act as intracellular signaling molecules. For example, when animal cells undergo programmed cell death, or apoptosis, phosphorylated inositol phospholipids act as binding sites that recruit specific proteins from the cytosol to the membrane. Phosphatidylserine, which is normally confined to the cytosolic monolayer of the plasma membrane lipid bilayer, rapidly translocates to the extracellular monolayer The phosphatidylserine exposed on the cell surface serves as a signal to induce neighboring cells, such as macrophages, to phagocytose and digest the dead cell.

Some functions of membrane phospholipids in cell signaling. (A) Extracellular signals can activate PI 3-kinase, which phosphorylates inositol phospholipids in the plasma membrane. Signaling molecules then bind to these phosphorylated lipids & are thus recruited to the membrane, where they can interact and help relay the signal into the cell. (B) Other extracellular signals activate phospholipases that cleave phospholipids. The lipid fragments then act as signaling molecules to relay the signal into the cell.

Freeze-fracturing splits the bilayer, showing proteins to be entirely within the lipid bilayer (integral/ transmembrane/ intrinsic), or projecting from the outer surface to varying degrees (peripheral/ extrinsic).

PROTEINS

1.Transmembrane, integral or intrinsic proteins extend through the lipid bilayer, with part of their mass on either side.  are amphiphatic molecules possessing one or more hydrophobic regions that exhibit an affinity for the hydrophobic interior of the lipid bilayer; hence these molecules are difficult to remove from membranes  They are notoriously difficult to crystallize; few have been studied in their entirety by x-ray crystallography.  DNA cloning and sequencing techniques, have revealed their amino acid sequences, and it is often possible to predict from an analysis of the protein's sequence which parts of the polypeptide chain extend across the lipid bilayer.  Proteins are capable of lateral movement within the bilayer; apical to basolateral movement is hindered by cell junctions, or their attachment to cytoskeletal elements.

2.Peripheral or extrinsic proteins function on only one side of the lipid bilayer; are often associated exclusively with either the lipid monolayer or a protein domain on that side.  Some of these are anchored to the cytosolic surface by an amphipathic helix that partitions into the cytosolic monolayer of the lipid bilayer through the hydrophobic face of the helix. For example, some of the proteins involved in intracellular signaling are bound to the cytosolic half of the plasma membrane by one or more covalently attached lipid groups.

Peripheral proteins of RBC plasma membrane are spectrin and ankyrin, and a protein called band. These proteins are bound to the inner surface of PM and form as a meshwork that support the PM and help maintain the shape of RBC. Gene mutations for these proteins result to RBC shape anomalies.

 How a membrane protein associates with the lipid bilayer reflects the function of the protein. Only transmembrane proteins (e.g. cell-surface receptors can function on both sides of the bilayer or transport molecules across it.

Various ways in which membrane proteins associate with the lipid bilayer. Some of these "single-pass" and "multipass" transmembrane proteins have a covalently attached fatty acid chain inserted in the cytosolic lipid monolayer (1). Most are thought to extend across the bilayer as (1) a single a helix, (2) as multiple a helices, (3) as a rolled-up sheet or a barrel. Others (5) are attached to the bilayer by a covalently attached lipid chain either a fatty acid chain or a prenyl group in the cytosolic monolayer, (6) via an oligosaccharide linker, or to phosphatidylinositol in the noncytosolic monolayer. (7, 8) Many proteins are attached to the membrane only by noncovalent interactions with other membrane proteins.

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 contribute to the glycocalyx in many cells. Note that all of the carbohydrate is on the noncytosolic surface of the membrane.

 It plays a role in the selective uptake of substances, recognition of self, a cellular ID system.  It confers an electrostatic charge on the cell surface, important in cell-to-cell adhesion. Disturbances in the cell coat and surface charge maybe responsible for the failure of cancer cells to adhere to one another. Histogenesis is achieved by modulating celladhesion molecules (CAMs) which are integral protein-carbohydrate (e.g., sialic acid) complexes. Antigens are located within the cell coat.

Human ABO bloodgroup antigens. These antigens are oligosaccharide chains covalently attached to glycolipids or glycoproteins in the plasma membrane. The terminal oligosaccharide sugars distinguish the 3 antigens. The presence or absence of the glycosyltransferases that add galactose (Gal) or Nacetylgalactosamine (GalNAc) to O antigen determine a persons blood type.

TRANSPORT MECHANISMS

1. Passive transport down an electrochemical gradient occurs spontaneously, either by simple diffusion through the lipid bilayer or by facilitated diffusion through channels and passive carriers.

A model of how a conformational change in a carrier protein could mediate the PASSIVE TRANSPORT of a solute. The carrier protein can exist in 2 conformational states: in state A, the binding sites for solute are exposed on the outside of the lipid bilayer; in state B, the same sites are exposed on the other side of the bilayer. The transition between the two states can occur randomly, completely reversible and does not depend on whether the solute binding site is occupied.

2. Active transport requires an input of metabolic energy and is always mediated by carriers that harvest metabolic energy to pump the solute against its electrochemical gradient. An electrochemical gradient combines the membrane potential and the concentration gradient, which can work additively to increase the driving force on an ion across the membrane or can work against each other.

The sodium-potassium pump: a specific case of active transport

http://highered.mcgrawhill.com/olc/dl/120068/bio03.swf

transports the solute in (or out) and the cotransported solute the opposite direction.

http://higher ed.mcgrawhill.com/olc/ dl/120068/bi o04.swf

transports the solute and a co-transported solute at the same time in the same direction.

http://higher ed.mcgrawhill.com/olc/ dl/120068/bi o04.swf

http://highered.mcgrawhill.com/olc/dl/120068/bio05.swf

Transcellular transport of glucose across an intestinal epithelial cell

Co-transport
Glucose is pumped into the cell through the apical domain of the membrane by a Na+ -powered glucose symport. Glucose passes out of the cell by passive transport mediated by a different glucose carrier protein in the basal and lateral membrane domains.

4.Signal-Reception- messenger molecules are membrane-bound or may pass through membrane channels to mediate communication among cells. G-proteins- activated by the binding of a 1st messenger molecule to a membrane receptor. The GTP-G protein exchanges act on membrane-bound enzyme effectors, which in turn activate a 2nd messenger. This triggers a cascade of molecular reactions that lead to changes in cell behavior (e.g., LH activates G5 adenylyl cyclase increased synthesis of estrogen and progesterone).

 The gating of ion channels. Different kinds of stimuli open ion channels. Mechanically gated channels often have cytoplasmic extensions that link the channel to the cytoskeleton.  Ion Channel Diseases:  Myasthenia Gravis Muscle weakness due to autoantibodies against the acetylcholine receptor  Cystic Fibrosis Defect in the Cl- channel CFTR leads to excessive phlegm and static infections

when transport is out of the cell.

http://highered .mcgrawhill.com/olc/dl/ 120068/bio02. swf