Dr.

Andrew Johan, 08179523380 Cell membrane = plasma membrane

Q: How do things enter and exit the cell? A: Molecules can enter a cell across the cell membrane by going through the PHOSPHOLIPID BILAYER or by using a special protein in the cell membrane. Cell membrane: 1. serves as a gateway and a barrier for the cell:control what enters and exits the cell. 2. is composed of phospholipids, proteins and carbohydrate 3. the heads of the phospholipids hydrophilic 4. the tails do not like to be with water (hydrophobic/lipophilic)

A. INTRACELLULAR FLUID (ICF)

This compartment constitutes two-thirds of total body water and provides the environment for the cell: (1) to make, store, and utilize energy; (2) to repair itself; (3) to replicate; and (4) to perform special functions.

B. EXTRACELLULAR FLUID (ECF)

This compartment contains about one-third of total body water and is distributed between the plasma and interstitial compartments. The extracellular fluid is a delivery system. It brings to the cells nutrients (eg, glucose, fatty acids, amino acids), oxygen, various ions and trace minerals, and a variety of regulatory molecules (hormones) that coordinate the functions of widely separated cells. Extracellular fluid removes CO 2,

waste, products, and toxic or detoxified materials from the cellular environment. ECF
Na+ 140 mmol/L K+ 4 mmol/L Ca2+ 2.5 mmol/L Mg2+ 1.5 mmol/L PO43 2 mmol/L CI 100 mmol/L Glucose 5.5 mmol/L Protein 2 g/dL

ICF
10 mmol/L 140 mmol/L 0.1 mol/L 30 mmol/L 60 mmol/L 4 mmol/L 01 mmol/L 16 g/dL

internal environment is rich in K+ and Mg2+, and phosphate is its major anion. Extracellular fluid is characterized by high Na+ and Ca2+ content, and Clis the major anion. Note also that the concentration of glucose is higher in extracellular fluid than in the cell, whereas the opposite is true for proteins. Membrane Structure and Composition

The table below shows some of the variation in protein, lipid and carbohydrate content of some membranes. membrane % protein % lipid % Carbohydrate myelin 18 79 3 human erythrocyte plasma membrane 49 43 8 mitochondrial inner membrane 79 24 0 amoeba plasma membrane 54 42 4 Q: Which of the following membranes contain highest amount of lipid ? All major lipids in membranes contain both hydrophobic and hydrophilic regions and are therefore termed amphipathic.

The major lipids in mammalian membranes are: phospholipids, glycosphingolipids, & cholesterol
The sugar derivative and alcohols that may be linked to the phosphate in phosphoglycerides are: inositol, choline, ethanolamine, and serine. phospholipids differ in the charge : Phospholipids that have no net electric charge at neutral pH: phosphatidylcholine and phosphatidylethanolamine phosphatidylglycerol and phosphatidylserine have a net negative charge.

Cholesterol in membranes

Transmembrane protein: 1. ion channel

2. protein transporter

3. receptor

4. enzyme

Membranes Are Asymmetric Structures This asymmetry can be partially attributed to: 1. the irregular distribution of proteins within the membranes, 2. the external location of the carbohydrates attached to membrane proteins, 3. inside-outside (transverse) asymmetry of the phospholipids. The choline-containing phospholipids (phosphatidylcholine and sphingomyelin) are located mainly in the outer molecular layer; the aminophospholipids (phosphatidylserine and phosphatidylethanolamine) are preferentially located in the inner leaflet.

PERIPHERAL PROTEINS lie on the surface of the membrane. Other proteins, called INTEGRAL PROTEINS, extend into and sometimes completely through the membrane.Proteins are the major functional molecules of membranes and consist of enzymes, pumps and channels, structural components, antigens, and receptors. Red blood cells (erythrocytes) have no nucleus or internal organelles, the plasma membrane is their only membrane, and it can be isolated without contamination by internal membranes (thus avoiding a serious problem encountered in plasma membrane preparations from other eukaryotic cell types.
Membrane Plasma membrane Endoplasmic reticulum Inner mitochondrial membrane Enzyme 5-Nucleotidase, Adenylyl cyclase,Na+-K+ ATPase Glucose-6-phosphatase ATP synthase

Passive Transport Diffusion (downhill transport) is movement of molecules or ions: only when the solute concentration at the starting point is higher than at the target site (unidirectional). In absence of a concentration gradienti.e., at equilibriumnet diffusion is zero because there is equal flux in both directions). The driving force of diffusion is, therefore, a concentration gradient . Example: When a layer of O2 gas is placed on water, the O2 quickly diffuses into the water along the initially high gas pressure gradient. As a result, the partial pressure of O2 (PO2) rises, and O2 can diffuse downward , for example, when O2 diffuses from the alveoli of the lungs into the bloodstream and when CO2 diffuses in the opposite direction.

Nonionic diffusion occurs when the uncharged form of a weak base (e.g., ammonia = NH3) NH4+ or acid (e.g., formic acid, HCOOH) passes through a membrane more readily than the charged form. In this case, the membrane would be more permeable to NH3 than to NH4+ NH NH3 3 4

The following factors affect net diffusion of a substance: (1) Its concentration gradient across the membrane. Solutes move from high to low concentration. (2) The electrical potential across the membrane. Solutes move toward the solution that has the opposite charge. The inside of the cell usually has a negative charge. (3) The permeability coefficient of the substance for the membrane. (4) The hydrostatic pressure gradient across the membrane. (5) Temperature. Increased temperature will increase particle motion and thus increase the frequency of collisions between external particles and the membrane. In addition, a multitude of channels exist in membranes that route the entry of ions into cells.

The membranes of nerve cells contain well-studied ion channels that are responsible for the action potentials generated across the membrane. One ion can regulate the activity of the channel of another ion. For example, a decrease of Ca2+ concentration in the extracellular fluid increases membrane permeability and increases the diffusion of Na+. This depolarizes the membrane and triggers nerve discharge. Facilitated diffusion and active transport share many features. Both appear to involve carrier proteins, and both show specificity for ions, sugars, and amino acids. Facilitated diffusion and active transport resemble a substrate-enzyme reaction except that no covalent interaction occurs. These points of resemblance are as follows: (1) There is a specific binding site for the solute. (2) The carrier is saturable, so it has a maximum rate of transport (Vmax). (3) There is a binding constant (Km) for the solute, and so the whole system has a Km. (4) Structurally similar competitive inhibitors block transport. Major differences are the following: (1) Facilitated diffusion can operate bidirectionally, whereas active transport is usually unidirectional. (2) Active transport always occurs against 5

an electrical or chemical gradient, and so it requires energy. The rate at which solutes enter a cell by facilitated diffusion is determined by the following factors: (1) The concentration gradient across the membrane. (2) The amount of carrier available. (3) The rapidity of the solute-carrier interaction. (4) The rapidity of the conformational change for both the loaded and the unloaded carrier.

specific solutes diffuse down electrochemical gradients across membranes more rapidly than might be expected from their size, charge, or partition coefficients. This facilitated diffusion exhibits properties distinct from those of simple diffusion. The rate of facilitated diffusion, a uniport system, can be saturated; ie, the number of sites involved in diffusion of the specific solutes appears finite. Many facilitated diffusion systems are stereospecific but, like simple diffusion, require no metabolic energy.

Hormones regulate facilitated diffusion by changing the number of transporters available. Insulin increases glucose transport by recruiting transporters from an intracellular reservoir. Insulin also enhances amino acid transport in liver and other tissues. Glucocorticoid hormones enhance transport of amino acids into liver, where the amino acids then serve as a substrate for gluconeogenesis.Growth hormone increases amino acid transport in all cells, and estrogens do this in the uterus. Valinomycin is a passive carrier for K+. It can bind or release K+ when it encounters the membrane surface. Valinomycin can catalyze net K+ transport because it can translocate either in the complexed or uncomplexed state. The direction of net flux depends on the electrochemical K+ gradient.

Property Carrier Mediated

Against Concentration Gradient

simple diffusion Facilitated Diffusion Active Transport + + + 6

Specificity Energy Expended

+ -

+ +

The erythrocyte membrane contains a Cl/HCO3 anion antiporter (AE1 protein) that facilitates transport of CO2 by the blood. Kadar klorida lebih tinggi pada eritrosit vena perifer dari pada vena pulmonalis SEBAB terjadi pertukaran antara ion Cl dan HCO3 Classes of Carrier Proteins: Uniport carriers mediate transport of a single solute. An example is the GLUT1 glucose carrier. Valinomycin is a passive carrier for K+. uniport system moves one type of molecule
bidirectionally.

Symport carriers bind two dissimilar solutes (substrates) and transport them together across a membrane. Transport of the two solutes is obligatorily coupled, solutes in the same direction. A gradient of one substrate, usually an ion, may drive uphill (against the gradient) transport of a co-substrate. SGLT: sodium glucose transporter.Glucose and Na+ bind to different sites on the
glucose transporter. Na+ moves into the cell down its electrochemical gradient and drags glucose with it Therefore, the greater the Na+ gradient, the more glucose enters; and if Na+ in extracellular fluid is low, glucose transport stops. this Na+-glucose symport is dependent on gradients generated by an Na +-K+ pump that maintains a low intracellular Na+ concentration.

Antiport (exchange diffusion) carriers exchange one solute for another across a membrane.

catalyzes 1:1 exchange of ADP for ATP across the inner mitochondrial membrane

Active transport. The Na+-K+ pump is oriented so that it pumps Na+ out of the cell and K+ into it. 1 ATP used for exporting 3 Na+ ions and importing 2 K+ ions. Crucial for maintaining resting membrane potential.

Intracellular Transport Lysosomes: Uptake of H+ ions from the cytosol and release of metabolites such as amino acids into the cytosol. Endoplasmic reticulum (ER): In addition to a translocator protein, the ER has two other proteins that transport Ca2+. Ca2+ can be pumped from the cytosol into the ER by a Ca2+-ATPase called SERCA (sarcoplasmic endoplasmic reticulum Ca2+-transporting ATPase). The resulting Ca2+ stores can be released into the cytosol via a Ca2+ channel (ryanodine receptor, RyR) in response to a triggering signal. Mitochondria: The outer membrane contains large pores called porins that render it permeable to small molecules (< 5 kDa). The inner membrane has high concentrations of specific carriers and enzymes. Enzyme complexes of the respiratory chain transfer electrons from high to low energy levels, thereby pumping H+ ions from the matrix space into the intermembrane space, resulting in the formation of an H+ ion gradient directed into the matrix. This not only drives ATP synthetase (ATP production, but also promotes the inflow of pyruvate and anorganic phosphate (symport). Ca2+ ions that regulate Ca2+-sensitive mitochondrial enzymes in muscle tissue can be pumped into the matrix space with ATP expenditure, thereby allowing the mitochondria to form a sort of Ca2+ buffer space for protection against dangerously high concentrations of Ca2+ in the cytosol. The inside negative membrane potential (caused by H+ release) drives the uptake of ADP in exchange for ATP (potential-driven transport). Transport between Adjacent Cells 8

Transport between adjacent cells occurs either via diffusion through the extracellular space (e.g., paracrine hormone effects) or through channel-like connecting structures (connexons) located within a so-called gap junction or nexus. A connexon is a hemichannel formed by six connexin molecules. One connexon docks with another connexon on an adjacent cell, thereby forming a common channel through which substances with molecular masses of up to around 1 kDa can pass. Since this applies not only for ions such as Ca2+, but also for a number of organic substances such as ATP, these types of cells are united to form a close electrical and metabolic unit (syncytium), as is present in the epithelium, many smooth muscles, the myocardium, and the glia of the central nervous system. Electric coupling permits the transfer of excitation, e.g., from excited muscle cells to their adjacent cells, making it possible to trigger awave of excitation across wide regions of an organ, such as the stomach, intestine, biliary tract, uterus, ureter, atrium, and ventricles of the heart. Certain neurons of the retina and CNS also communicate in this manner (electric synapses).

ion channels: Ion channels can be highly selective for particular ions. For example, some channels allow the flow of K+ very effectively but do not allow appreciable levels of Na+ to cross the membrane. Other channels transport positively charged ions (cations), but block the flow of negatively charged ions (anions). Ion channels exist in open and closed states. These channels undergo transitions from the closed state, incapable of supporting ion transport, to the open state, through which ions can flow. Transitions between the open and the closed states are regulated. Ion channels are divided into two classes: ligand-gated channels and voltage-gated channels. Ligand-gated channels open and close in response to the binding of specific chemicals, whereas voltage-gated channels open and close in response to the electrical potential across the membrane in which they are found. The ligand-gated channel; the acetylcholine receptor channel, which 9

communicates the nerve impulse between certain neurons; Snake neurotoxins such as -bungarotoxin (from the venom of a Formosan snake) and cobratoxin block the transmission of impulses between nerve and muscle. These small (7-kd) basic proteins bind specifically and very tightly to acetylcholine receptors.Acetylcholine opens a single kind of cation channel, which is almost equally permeable to Na+and K+. Voltage-gated channel contohnya: Na+ channels. Depolarization of the membrane beyond the threshold level leads to an opening of Na+ channels. Sodium ions begin to flow into the cell because of the large electrochemical gradient across the plasma membrane. The entry of Na+ further depolarizes the membrane, and so more gates for Na+ are opened. This positive feedback between depolarization and Na+ entry leads to a very rapid and large change in membrane potential, from about -60 mV to +30 mV in a millisecond. A specific neurotoxin. Tetrodotoxin, an organic compound isolated from the puffer fish, binds to sodium channels with great avidity. block movement of Na+ through Na+ channel.The lethal dose of this poison for an adult human being is about 10 ng.

A potent neurotoxin such as Veratridine and batrachotoxin affects the nervous system by causing depolarization of nerve and muscle fibres due to increased sodium ion permeability of the excitable cell membrane.keep Na+ channel permanently open. In 1785, William Withering, a British physician, heard tales of an elderly woman, known as "the old woman of Shropshire," who cured people of "dropsy" (which today would be recognized as congestive heart failure) with an extract of foxglove. Withering conducted the first scientific study of the effects of foxglove on congestive heart failure and documented its effectiveness. ouabain and digoxin increase the force of heart muscle contraction and are widely used in the treatment of congestive heart failure. The primary effect of these drugs is to inhibit the Na+/K+ ATPase, thereby raising the intracellular Na+ concentration (and lowering intracellular K+). Because the Na+/Ca2+ antiporter functions less efficiently with a lower Na+ concentration gradient, fewer Ca2+ ions are exported and the intracellular Ca2+ concentration increases. This increase causes the muscle to contract more strongly. Inhibit Na+/K+ ATPase --> raising intracellular Na+ concentration --> Na+/Ca2+ antiporter functions less efficiently --> fewer Ca2+ ions are exported --> intracellular Ca2+ concentration increases --> causes the muscle to contract more strongly. In cardiac muscle cells, the export of Ca2+ is coupled to the import of Na+ by a cation antiporter, which transports 3 Na+ ions inward for each Ca2+ ion exported. Open states of channels often spontaneously convert into inactivated states. determine the 10

duration of ion flow such as those lining the small intestine and the kidney tubules, need to concentrate glucose against a very large concentration gradient. Such cells utilize a two Na+/one-glucose symporter; a protein that couples transmembrane movement of one glucose molecule to the transport of two Na+ ions:
Properties of ion channels
* They are composed of transmembrane protein subunits. Most are highly selective for one ion; a few are nonselective. They allow impermeable ions to cross membranes at rates approaching diffusion limits. They can permit ion fluxes of 106107/s. Their activities are regulated. The two main types are voltage-gated and ligand-gated. They are usually highly conserved across species. Most cells have a variety of Na+, K+, Ca2+, and CIchannels. Their activities are affected by certain drugs.

In cardiac muscle cells, Na+/Ca2+ antiporter plays the principal role in maintaining a low concentration of Ca2+ in the cytosol. The reaction of this cation antiporter can be written:

The drugs ouabain and digoxin increase the force of heart muscle contraction and are widely used in the treatment of congestive heart failure. The primary effect of these drugs is to inhibit the Na+/K+ ATPase, thereby raising the intracellular Na+ concentration (and lowering intracellular K+). Because the Na+/Ca2+ antiporter functions less efficiently with a lower Na+ concentration gradient, fewer Ca2+ ions are exported and the intracellular Ca2+ concentration increases. This increase causes the muscle to contract more strongly. Neurotoxin Tetradotoxin and saxitoxin block movement of Na+ through Na+ channel. Botulinus toxin inhibits acetylcholine release. Veratridine and batrachotoxin keep Na+ channel permanently open.

SUMMARY Membranes are complex structures composed of lipids, carbohydrates, and proteins. The basic structure of all membranes is the lipid bilayer. This bilayer is formed by two sheets of phospholipids in which the hydrophilic polar head groups are directed away from each other and are exposed to the aqueous environment on the outer and inner surfaces of the membrane. The hydrophobic nonpolar tails of these molecules are oriented toward each other, in the direction of the center of the membrane. 11

 Membrane proteins are classified as integral if they are firmly embedded in the bilayer and as peripheral if they are loosely attached to the outer or inner surface. Certain molecules freely diffuse across membranes, but the movement of others is restricted because of size, charge, or solubility. Various passive and active mechanisms are employed to maintain gradients of such molecules across different membranes. Certain solutes, eg, glucose, enter cells by facilitated diffusion, along a downhill gradient from high to low concentration. Specific carrier molecules, or transporters, are involved in such processes. Ligand- or voltage-gated ion channels are often employed to move charged molecules (Na+, K+, Ca2+, etc) across membranes. Large molecules can enter or leave cells through mechanisms such as endocytosis or exocytosis. These processes often require binding of the molecule to a receptor, which affords specificity to the process. Receptors may be integral components of membranes (particularly the plasma membrane). The interaction of a ligand with its receptor may not involve the movement of either into the cell, but the interaction results in the generation of a signal that influences intracellular processes (transmembrane signaling). Mutations that affect the structure of membrane proteins (receptors, transporters, ion channels, enzymes, and structural proteins) may cause diseases; examples include cystic fibrosis and familial hypercholesterolemia.

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