BIOL2123:

Module 3 CELL MEMBRANE:

It is now a routine task to determine the exact order in which individual subunits have been linked together in polynucleotides (DNA) and polypeptides (proteins). However, it remains difficult to determine the arrangement of monomers in a polysaccharide. Explain why this is the case. DNA has a predictable patterns with 4 nucleotides Polypeptides have 20 amino acids. Some polysaccharides are branched do not know whether you are sequencing from main chain or branches. Complex structural organisation. Alberts et al. Essential Cell Biology Chapter 11 pp.363-385 Chapter 12 pp. 387-401 Chapter 15 pp.510-524 SINGER & NICOLSON MODEL: 1972 revised model Proteins dispersed and individually inserted into phospholipid bilayer Only hydrophilic regions protruding are exposed to water Membrane is a mosaic of protein in a fluid bilayer of phospholipid Fluid mosaic model Fluid: not a rigid structure phospholipids can move within structure Mosaic: mixture of compounds proteins embedded, phospholipids Lipid bilayer is a 2D fluid Fluidity and flexibility Phospholipids are mobile within a layer Hydrocarbon chains able to rotate Fatty acid tail: More unsaturated bonds leads to more fluid membrane structure Chain length important Single bonds tightly packed Electron spin resonances rotate on axis. Diffuse in 2 dimensions on membrane Dynamic structure

MEMBRANE STRUCTURE: Protein-studded, fatty film Cannot be seen directly in light microscope. Serve as barrier to prevent content of cell from escaping and mixing with surrounding medium. Eukaryotic cell: consist of internal membrane. Enclose intracellular compartments. Endoplasmic reticulum, Golgi apparatus, mitochondria, chloroplast. Internal membranes and external membranes have subtle differences in composition. 5 nm (50 atoms) thickness Selective barrier selectively permeable (external and internal environment) Diffuse from high concentration to low concentration

Ion channels, selective complex channels Facilitates exchange and has dynamic structure Protein (function) and carbohydrate (function) Cell signal recognition Universal and specific functions depending on cell type Receptor proteins: receive information/signals from the environment. Transport proteins: Import/export small molecules Flexibility: enables capacity for movement and expansion. Enlarge in area adding new membrane without losing continuity Can deform without tearing. Small size need to concentrate certain metabolites in small area Bacteria: plasma membrane only in cell membrane Eukaryote: internal membrane and cell membrane

CELL COMMUNICATION: Import and export molecules Cell growth and motility. 1. Receptor proteins enable cell to receive signals form environment. 2. Transport protein in membrane enable import and export of small molecules. 3. Flexibility of membrane and capacity for expansion allow cell to grow, change, shape and move. DOUBLE MEMBRANE ORGANELLES: Nucleus and mitochondria Chloroplast. Fatty acids are hydrophobic Phospholipids amphipathic AMPHIPATHIC: Crucial in driving lipid molecules to assemble into bilayers in aqueous solution. Two conflicting forces HYDROPHILIC: Charged group or uncharged polar group. Form electrostatic attractions or hydrogen bonds with water. HYDROPHOBIC: Most/all atoms uncharged and nonpolar. Cannot form favourable interactions with water molecules. Force adjacent water molecule to reorganise to cage-like structure highly ordered. Cage-like structure: Formation requires free energy. Energy cost minimised when hydrophobic molecules cluster together limit contact with water. Coalesce into single large drop in water.

LIPID BILAYER:
Two closely apposed sheets form lipid bilayer. Fundamental structure of all cell membrane

Serve as permeable barrier to most water-soluble molecules. Membrane lipids form bilayers in water phospholipid bilayer The lipid bilayer is a two-dimensional fluid The fluidity of a lipid bilayer depends on its composition Phospholipid on outer part different from inner part. Lipid asymmetry is preserved during membrane transport Cell will move towards most energetically favourable position. Amphipathic molecules self-sealing. Tears in bilayer energetically unfavourable. Spontaneously rearrange eliminate free edge (tear). Bend and seal form boundary around closed space. Small tear: rearrangement exclude water molecules repair of bilayer and restore single continuous sheet. Large tear: sheet folds itself and break into separate closed vesicles.

Movement of lipid molecules Variety of movements - frequent and rapid. Flip-flops: tumble from one monolayer to another. Without proteins to facilitate process. Random thermal motions: lipid molecule continuously exchange places with neighbour (same monolayer). Rapid lateral diffusion of lipid molecules Individual lipid molecules flex hydrocarbon tail and rotate rapidly about long axis. Transfers rarely occur spontaneously catalysed by scramblases. Scramblases remove randomly selected phospholipids from one half on bilayer and insert them to another. FLUIDITY LIPID BILAYER: Temperature depends on composition length and number of double bonds. More viscous/less fluid: closer and regular packing of tails. Shorter chain: reduce tendency of hydrocarbons to interact increase fluidity Double bond: large portion of unsaturated hydrocarbon tails more fluid than lower proportions. Animals: membrane fluidity modulated by cholesterol. Membrane held together by hydrophobic interactions

Most lipids drift around randomly in the plane of the membrane Rarely do phospholipids flip from one side to another Phospholipids move at 2 m per second Proteins move much slower PHOSPHATIDYLCHOLINE: Small molecule choline attached to phosphate group (hydrophilic head) Most common phospholipid. Phosphatidyl: phosphate-glycerolfatty acid portion. TRIACYLGLYCEROL: Main constituents of animal fats and plant oils. Entirely hydrophobic. Store energy Fatty acid and glycerol. PHOSPHOLIPID: Different phospholipids have different polar group. Amphipathic Most abundant lipid in cell. Phosphate (hydrophilic head) linked to pair of hydrophobic tails.

CHOLESTEROL: Stabilise membrane stiffen and less flexible/permeable. o At 37 C stabilises/restrains phospholipids At lower temperatures hinders close packing of phospholipids Membranes do not solidify Amphipathic Molecules short and rigid. Cholesterol fits into gaps between phospholipid molecules in lipid bilayer.

MEMBRANE ASSEMBLY:
EUKARYOTIC CELLS: Phospholipids manufactured by enzymes bound to cytosolic surface of ER. 1. Use free fatty acid as substrate. 2. Enzyme deposit newly made phospholipids exclusively in cytosolic (inside) half of bilayer. Scramblases redistribute phospholipid. Newly made phospholipid redistributed equally between each monolayer of ER membrane. Newly assembled ER Remain in ER Supply fresh membrane to other compartments.

Membrane continually pinch off ER to small vesicles fuse with other membranes (ie. Golgi apparatus) plasma membrane. FUNCTION: Remove randomly selected phospholipids from one half on bilayer and insert them to another. Remove specific phospholipids from bilayer facing exterior and flip them to monolayer facing cytosol. Initiate and maintain asymmetrical arrangement of phospholipid. LOCATION: Endoplasmic reticulum.

ENZYME: Scramblases

Flippases

Golgi apparatus.

MEMBRANE SIDENESS: Membranes have a distinct inside and outside 2 halves of bilayer striking different sets of phospholipids. Lipid layers may be of different composition Proteins are of different composition Carbohydrates restricted to the outside Glycoproteins and glycolipid on present on surface of membrane Typical negative charge on inside of membrane Charged different on both sides electrical and chemical gradient. Constant material exchange Orientation between organelle and plasma membrane mirrored (symmetrical) due to vesicle movement. Exocytosis/endocytosis. Membrane fused inverted/opened

Cytosolic monolayer faces cytosol. Noncytosolic monolayer exposed to cell exterior Lumen: interior space of organelle.

Lipids (glycolipids) show most dramatic lopsided distribution. Sugar groups face exterior form continuous coat of carbohydrate: surround and protect animal cells. Located mainly in plasma membrane. Glycolipids acquire sugar groups in Golgi apparatus. Sugars added only to lipid molecules in noncytosolic half of bilayer. Glycomolecule trapped in monolayer no flippase to transfer glycolipids to cytosolic side.

FLUID MOSAIC: Membranes are a collage of different proteins embedded in fluid phospholipid matrix. Proteins determine characteristics of membrane More than 50 different kinds of proteins have been found in membranes of red blood cells. Cell membrane inside face: cytosolic monolayer face cytosol. Cell membrane outside face: noncytosolic monolayer exposed to cell exterior or interior space (lumen) or organelle. Sugar groups form continuous coat of carbohydrate surround and protect animal cell. Lopsided distribution (lipids) glycolipids. Located mainly in plasma membrane and noncytosolic half of bilayer. Acquire sugar group in Golgi apparatus.

1.

2.

3. 4.

Membrane proteins extended through bilayer part of their mass on either side. Transmembrane proteins amphipathic. Hydrophobic regions lie in interior of bilayer against hydrophobic tails of lipid molecule. Hydrophilic regions exposed to aqueous environment on either side of membrane. Other membrane proteins located almost entirely in cytosol. Associated with cytosolic half of lipid bilayer. Amphipathic alpha helix exposed on surface of protein. Some proteins lie entirely outside bilayer Attached to membrane only by one or more covalently attached lipid groups. Other proteins bound indirectly to one or other face of membrane. Held in place only by interactions with other membrane proteins.

MEMBRANE PROTEINS:
Membrane proteins membrane function. Lipid molecules much smaller than protein. Membrane proteins associate with lipid bilayer in various ways Make 50% of mass Polypeptide chain usually cross bilayer as Helix. Membrane protein solubilised in detergents and purified. Complete structure is known for relatively few membrane proteins Plasma membrane reinforced by cell cortex. Cells can restrict movement of membrane proteins. Cell surface coated with carbohydrate. FUNCTIONS: Transporter/ion channels Signal recognition/single transduction Receptor on membrane for uptake of ion, hormones Uptake in organised fashion Chemical signal perceived at membrane Change within cell is affected Intercellular joining cell adhesion Cell recognition Associated with carbohydrate component Attachment to cytoskeleton and extracellular matrix Receptors Enzymes maintain asymmetric arrangement. PROTIEN EXAMPLE: Na+ pump SPECIFIC FUNCTION: Actively pumps Na+ out of cells and K+ in. Allows K+ ions to leave cell, thereby having a major influence on cell excitability. Link intracellular actin filaments to extracellular matrix proteins. Binds extracellular PDGF and, as a consequence, generates intracellular signals that cause the cell to grow and divide. Catalyzes the production of the small intracellular signalling molecule of cyclic AMP in response to extracellular signals.

FUNCTIONAL CLASS: Transporters

Ion channels

K+ leak channel.

Anchors Receptors

Integrins Platelet-derived growth factor (PDGF) receptor.

Enzymes:

Adenylyl cyclase:

Enzymes orientated in such that sugars are added only to lipid molecules in noncytosolic half of bilayer. Once a glycolipid molecule has been created in this way, it remains trapped in this monolayer, as there are no flippases that transfer glycolipids to the cytosolic side. Thus, when a glycolipid molecule is finally delivered to the plasma membrane, it displays its sugar to the exterior of the cell.

(A) Transmembrane proteins can extend across the bilayer as a single helix, as multiple helices, or as a rolled-up sheet (called a barrel). (B) Some membrane proteins are anchored to the cytosolic surface by an amphipathic helix. (C) Others are attached to either side of the bilayer solely by a covalent attachment to a lipid molecule ( red zigzag lines). (D) Finally, many proteins are attached to the membrane only by relatively weak, non-covalent interactions with other membrane proteins. Side chain of amino acid hydrophobic Proteins attached directly to lipid bilayer transmembrane (associated with lipid monolayer) or lipid-linked (can be removed only by disrupting bilayer with detergents).

HELIX
Polypeptide chain usually crosses the lipid bilayer as an helix. Most common form in which polypeptide chain crosses lipid bilayer. Signal protein on outside of cell. Signal transductor in cytosol. Portions of transmembrane protein located on either side of the lipid bilayer are connected by specialized membrane-spanning segments of polypeptide chain. Run through hydrophobic environment of interior of lipid bilayer composed largely of amino acids with hydrophobic side chains. Side chains cannot form favourable interactions with water molecules prefer to interact with hydrophobic tails of lipid molecules (no water present). Hydrogen-bonds (in backbone) maximized if polypeptide chain forms regular helix. Many transmembrane proteins polypeptide chain crosses membrane only once. Receptors: single-pass transmembrane proteins Channels: proteins form aqueous pores across lipid bilayer to allow small, water-soluble molecules to cross membrane. Cannot be formed by proteins with single transmembrane helix. Consist of series of helices that cross bilayer a number of times multipass transmembrane proteins. Multipass transmembrane protein one or more membrane-spanning regions amphipathic. Formed from helices with amphipathic side chains.

SHEET/BARREL:
Rolled in cylinder keg-like structure: barrel. Hydrophilic inside side chains. Hydrophobic outer layer. Eg. PORIN proteins: large, water-filled pores in mitochondria and some bacteria outer membranes.

Allows passage of small nutrients, metabolites and inorganic ions across outer membrane. Prevent unwanted larger molecules from crossing.

PROTEINS AND DETERGENTS: Membrane proteins built to operate in environment partly aqueous and fatty. Individual proteins must be separated from all other cell proteins before detailed study solubilized in detergents. Solubilized with agents that destroy lipid bilayer disrupt hydrophobic associations. Disruptive agents: detergents. DETERGENTS: Small, amphipathic, lipid-like molecules. Single hydrophobic tail cone shaped. Aggregate into micelles. Hydrophobic ends interact with membrane-spanning hydrophobic regions of transmembrane proteins and hydrophobic tails of phospholipid molecules.

BACTERIORHODOPSIN: Small protein (250 amino acids) in plasma membrane of archaeans. Acts as membrane transport protein pump H+ (protons) out of cell. Pumping requires energy sunlight. Energy changes protein shape protein embedded in lipid bilayer undergo series of small conformational changes. Overall movement one H+ from inside to outside of cell. Energy generates energy gradient of H+ across plasma membrane. Bacteriorhodopsin is a pump protein a class of transmembrane proteins that actively moves small organic molecules and inorganic ions into and out of cells.

CELL CORTEX
Cell membranes strengthened and supported by framework of proteins attached to membrane via transmembrane proteins. Plants, yeas and bacteria: rigid cell wall meshwork of proteins, sugars and other macromolecules that encase plasma membrane. Shape and mechanical properties. Animal cells: Meshwork of fibrous proteins CELL CORTEX (attached to underside of membrane) Stabilise plasma membrane.

10

Cortex allow cell to selectively take up materials from environment, change shape, and move. Use cortex to restrain diffusion of proteins within plasma membrane.

Dimers linked to form longer tetramers. Tetramers (and actin molecules) linked into a mesh. Mesh attach to plasma membrane bind at least 2 types of attachment molecules.

DIMERIC PROTEIN SPECTRIN: Main component of red blood cell. Spectrin meshwork connected to membrane through intracellular attachment proteins that link spectrin to specific transmembrane proteins. Give structure. RESTRICTION OF PROTEIN MOVEMENT. Proteins can move freely within plane of lipid bilayer lateral diffusion. Membrane domains: confining particular proteins to localized areas within bilayer membrane, thereby creating functionally specialised regions on cell or organelle surface. Membrane proteins can be tethered to structures outside cell.

Proteins can diffuse laterally in their own membrane domains but are prevented from entering the other domain by a TIGHT JUNCTION (specialized cell junction). Basal lamina mat of extracellular matrix that supports all epithelial sheets. Apical surface: surface that faces the cytoplasm. Specialised functional proteins form continuous belt around cell where cell contacts is neighbours create seal between adjacent membrane.

11

INTEGRAL MEMBRANE PROTEINS: Generally transmembrane protein Only removed by disrupting bilayer with detergents. Hydrophobic regions completely span the membrane This region consists of one or more stretches of non-polar amino acids These are usually coiled into alpha helices Hydrophilic regions are exposed to aqueous solutions on either side of the membrane PERIPHERAL MEMBRANE PROTEINS: Not embedded in membrane. Gentle extraction interfere with protein-protein interactions leave lipid bilayer intact. Appendages loosely bound Often attached to integral proteins cytoplasmic side Some attached to cytoskeleton exterior Attached to extracellular matrix Strengthen animal cell

FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING (FRAP):

Uniformly labelling components of cell membrane (lipids, proteins) with fluorescent marker. Incubate living cells with fluorescent antibody or covantly attach fluorescent protein to membrane protein or interest Green fluorescent protein (GFP). Use recombinant DNA techniques. Small patch of membrane irradiated with intense pulse of light from sharply focused laser beam. Treatment irreversibly bleaches fluorescence from labelled proteins. Observe with fluorescent microscope Certain section bleached with laser saturate fluorescent. Diffuse with neighbour mix bleached and unbleached. Observe quick recovery Show that proteins on membrane very motile. If no recovery is seen protein not mobile in that area. Fluorescence recovery: direct measure of rate at which protein molecules can diffuse within the membrane.

12

CARBOHYDRATES ON CELL SURFACE:

Cell recognition Sugars covalently bonded to lipids in plasma membrane. Tissue recognition and rejection by immune system Due to surface carbohydrates Oligosaccharides: Short sugar chains. Less than 15 sugars long Some covalently attached to lipids. Glycoprotein oligosaccharides linked to proteins. Proteoglycans: Membrane protein contains one or more long polysaccharide chains. All carbohydrate on glycoproteins, proteoglycans, and glycoplipids is located on the outside of the plasma membrane forms sugar coating called GLYCOCALYX/carbohydrate layer.

GLYCOCALYX Protect cell surface from mechanical damage. Oligosaccharides and polysaccharides absorb water give slimy surface: help motile cells move through narrow spaces.

Positioned on exterior side of membrane Carbohydrate attached to Golgi apparatus tails modified in ER. Membrane lipids are amphipathic molecules most of which spontaneously form bilayers. Lipid bilayer is a 2D fluid The fluidity of a lipid bilayer depends on its composition The lipid bilayer is asymmetrical Glycolipids are found on the surface of all plasma membranes covalent bond Three major classes of membrane lipid molecules Lipid compositions of the inner and outer monolayers are different Membrane proteins can be associated with the lipid bilayer in various ways Electrical, noncovalent, covalent Most transmembrane proteins cross the lipid bilayer as an -helix Secondary coil structure. Bacteriorhodopsin is a proton pump that traverses the lipid bilayer as seven -helices Porins are pore-forming transmembrane proteins that cross the lipid bilayer as a barrel

13

Membrane proteins often function as large complexes Many membrane proteins diffuse in the plane of the membrane Cells can confine proteins and lipids to specific domains within a membrane The cell surface is coated with sugar residues Glycolipids or proteins.

14

MEMBRANE TRANSPORT:
Transfer depend on specialised membrane transport proteins Span lipid bilayer and provide private passageways across membrane for select substances. Transporters: shift small organic molecules or inorganic compounds by changing shape. Channels: form tiny hydrophilic pores across membranes through which such substances can pass by diffusion. Ion channels: Most channels only permit passage of inorganic ions. Movement create powerful electric force/voltage. Voltage differences enable nerve cell communication. Movement accelerated by facilitated transport. PERMEABLE: Small hydrophobic molecules: O2, CO2, N2, benzene Small, uncharged polar molecules: H2O, glycerol, ethanol IMPERMEABLE: Larger, uncharged polar molecules: Amino acids, glucose, nucleosides Ions (charged): H+, Na+. HCO3-, K+, Ca2+, Cl-, Mg2+ Excluded by size and charge.

LIPID BILAYERS: Lipid bilayers impermeable to ions and most uncharged polar molecules. Smaller molecule and more hydrophobic/nonpolar more rapidly diffuse. Polar/water-soluble require aid of membrane transport proteins. Small nonpolar molecules Uncharged polar molecules (Uneven distribution of electric charge): Charged molecules: Dissolve readily in lipid bilayers. Rapidly diffuse across. Diffuse readily if small enough. Larger uncharged polar molecules cross hardly at all (glucose) Lipid bilayer highly impermeable to all charged molecules. Including inorganic molecules. Inhibit entry to inner, hydrocarbon phase of bilayer. Oxygen, carbon dioxide. Water, glycerol.

IONS:
Most important inorganic ions: Na+, K+, Ca2+, Cl- and H+. Na+ Most plentiful +ive ion outsdie cell. K+ Most plentiful +ive ion inside cell. Quantity of positive charge inside cell and surrounding fluid must be balanced by an almost exact quantity of negative charge. Na+ balanced by Cl- outside cell. K+ balanced by ively charged organic and inorganic ions (anions). Membrane potential: electrical imbalances generate voltage difference across membrane. Cell unstimulated: exchange of anions and cations across the membrane precisely balanced. Resting state potential: steady-state conditions hold steady but not at 0. Animals: -20 and -200 mV. Value negative interior of cell more negatively charged than exterior. Membrane potential allows cells to power transport of certain metabolites and enable communication.

15

CLASSES OF MEMBRANE TRANSPORT

Each type of cell membrane has own characteristic set of transport proteins determine which solutes can pass. Traverse lipid bilayer form continuous protein-lined pathway allow selected small, hydrophilic molecules to cross without coming into direct contact with hydrophobic interior of lipid bilayer. Two main classes: Transporters Channels CHANNELS: Based on size and electric charge. Channel open: any ion/molecule small enough with appropriate charge pass. TRANSPORTER: Based on specific binding site. Bind with solutes with great specificity (enzyme bind to substrate).

ACTIVE & PASSIVE TRANSPORT: PASSIVE TRANSPORT: Molecules spontaneously flow downhill high concentration to low concentration (provided that pathway exist). No additional driving force no expenditure of energy by transport protein. Solutes move in both directions across membrane reach equilibrium. All channels and many transporters acts ads conduits for passive transport. ACTIVE TRANSPORT: Move solutes against concentration gradient. Transport protein requires an input of energy. Carried out by pumps harness energy source to power transport process. Energy from ATP hydrolysis, transmembrane ion gradient, sunlight.

PASSIVE TRANSPORT OF CHARGED SOLUTES: Uncharged molecule direction of passive transport determined by concentration gradient.

16

Membrane potential exerts force on any molecules that carries electric charge. Cytosolic side: negative potential relative to extracellular side. Membrane potential tends to pull positively charged solutes into cell Drive negatively charged solutes out. Charged solutes move down concentration gradient. Net force driving charged solute across a cell membrane composite of 2 forces Force due to concentration gradient Force due to membrane potential.

Electrochemical gradient: net driving force Determine direction of solute flow across membrane by passive transport. Na+: higher concentration outside cells enters a cell. K+: present at much higher concentrations inside cells than outside. Small electrochemical gradient across resting plasma membrane little net movement of K+ across membrane even when K+ channels open. PASSIVE MOVEMENT OF WATER: Water molecules small and uncharged. Diffuse directly but slowly. AQUAPORINS: specialized channel proteins in plasma membrane that facilitates in water flow. Increase rate of water flow across membrane. Osmolarity: total concentration of solute particles inside the cell. Osmolarity inside cell exceeds outside of cell osmotic gradient pull water into cell. Movement of water down concentration gradient OSMOSIS.

Cytoplasm: resists osmotic swelling. Contractile vacuoles: eliminate excess water by periodically discharging contents. Cell walls: tolerate large osmotic difference across plasma membrane. Turgor pressure: osmotic swelling pressure. Keep plant cell walls tense stems/leaves rigid and extended.

TRANSPORTERS: Responsible for most movement of small, water-soluble, organic molecules and inorganic ions. Highly selective often transfer one type of molecule. When molecule uncharged, electrochemical gradient it zero. Direction of movement depends on concentration gradient alone. Once receptors bind - protein switches conformation spontaneously and randomly. Passive transporters play no part in determining direction of transport only highly selective.

17

PUMPS & ACTIVE TRANSPORT:

ATP-driven pumps Coupled pumps Hydrolyze ATP to drive uphill transport. Link uphill transport of one solute across membrane to downhill transport of another. Mainly in bacterial cells. Use energy from sunlight to drive uphill transport.

Light-driven pumps

Na+ ATP PUMP: NA+-K+ ATPase or NA+-K+ pump: Na+ pump uses energy from ATP expel Na+ bring K+. ATP hydrolysis series of protein conformational changes that drive Na+/K+ ion exchange. Phosphate group removed from ATP transferred to pump. Cyclic and depends on previous steps to operate.

Ceaselessly expelling Na+ constantly entering cell through transporter and ion channels. Pump keeps Na+ concentration low and K+ concentration high. Steep gradient concentration create large Na+ electrochemical gradient pull Na+ back into cell.

Ca2+ ATP PUMP: Keep cytosolic Ca2+ concentration low. Less plentiful than Na+/ Ca2+ bind tightly to proteins alter activities/trigger cell process. Lower background concentration of Ca2+ more sensitive to increase in cytosolic Ca2+. Eukaryotic cells maintain very low concentration of free Ca2+ in cytosol. But much greater extracellular concentration. ATP-driven CA2+ pumps actively pump Ca2+ out of cytosol.

18

Lead to huge concentration difference (electrochemical gradient). Return to original conformation without requirement for binding and transporting a second ion. Difference to Na+ pump ^

COUPLED PUMPS:
Downhill movement of first solute provide energy for second uphill transport. Coupled pumps active transporters. Symport: pump moves both solute in same direction across membrane. Antiport: pump moves solutes in opposite directions. Uniport: pump moves only one type of solute across membrane.

Na+ GRADIENT COUPLED PUMPS: In animals. Two molecules must be present for coupled transport to occur Na+ and glucose When Na+ moves, glucose dragged with it. Binding of Na+ and glucose cooperative. Binding of two molecules cooperative binding of one enhances the binding of another. Two types of molecules (glucose) transporters segregated in proper domains by diffusion barrier formed by tight junction around apex of cell. Use influx of Na+ to pump H+ out of cell.

19

H+ GRADIENT COUPLED PUMPS: Plants cells, bacteria, and fungi (yeast). H+ pump substituted for Na+ pump. Pump H+ out of cell. H+ gradient created by activity of light-driven H+ pumps (bacteriorhodopsin) Used to maintain pH levels.

ION CHANNELS:
Ion selectivity permit some inorganic ions to pass. Depend on diameter, shape, distribution of charge amino acids. Each ion in aqueous solution contain water molecule shell must be shed in order to pass through. Pass through single file ions come into contact with channel wall Ion channels not continuously open. Channels gated: specific stimulus triggers them to switch between closed and open state change in conformation. (Unlike transporters) Open ion channel does not need to undergo conformational changes. Able to reach maximum rate of transport.

20

Cannot carry out active transport. Mainly Na+, K+, Ca2+ or ClIon channel opens ion flow through move rapidly down electrochemical gradient change membrane potential.

SPECIFIC ION MEMBRANE PERMEABILITY:

K+ actively imported into cell by Na+ pump. K+ leak channels: control movement of K+. Open: allow K+ to move freely tendency to flow out of cell down gradient. In resting state: K+ leak channels open Membrane much more permeable to K+. Equilibrium condition established keep K+ inside cell just strong enough to counteract tendency of K+ to move down concentration gradient concentration gradient of K+ is 0. Even though higher concentration inside cell. Resting membrane potential: flow of positive and negative ion across membrane precisely balanced no further difference in charge accumulates. Cell stimulated: ion channels open change membranes permeability to ions. Membrane potential at any time depends on both state of membranes ion channel and ion concentrations on either side of membrane. Bulk changes in ion concentrations cannot occur quickly enough to drive rapid changes in membrane potential associated with electrical signalling. Rapid opening and closing of ion channels. Net negative charge inside cell Net positive charge outside cell.

21

ADDITIONAL NOTES:
Concentration Gradients and Electrical Forces Drive Passive Transport Active Transport Moves Solutes Against Their Electrochemical Gradients + Animal Cells Use the Energy of ATP Hydrolysis to Pump Out Na + + The Na -K Pump Is Driven by the Transient Addition of a Phosphate Group + + The Na -K Pump Helps Maintain the Osmotic Balance of Animal Cells 2+ 2+ Intracellular Ca Concentrations Are Kept Low by Ca Pumps Coupled Transporters Exploit Gradients to Take Up Nutrients Actively + H Gradients Are Used to Drive Membrane Transport in Plants, Fungi, and Bacteria

Transport molecules move molecules against concentration gradient. Numerous different interactions. ABC cassettes transport phosphate group. ATP ADP +Pi Conformational change in protein allows release of protein. Use a form of energy to drive process against concentration gradient. Well characterized in animal cell membranes. Heterodimer protein 2 different polypeptide subunits. 2 different dimers. Different binding sites. Drive Na+ against concentration gradient. Dock K+ conformational change uptake of K+ into cell. Electrochemical gradient of Na+ and K+ Cl- damaging to cell.

Plants cell maintain osmotic balance through cell walls Protozoans: maintain balance by discharging contractile vacuole. Rate based on solute concentration in cell. Calcium also stored in mitochondria

22

Drive glucose against concentration gradient Anti-porter example of transporter. Cell can have different distribution of cell components. Tightly packed adhesion junctions. Generate barrier no ready movement Increase surface area more processes. Spend a lot of energy to pump NA+ out Use energy of diffusion to drive uptake of glucose. Glucose out uniport transport no energy required. Diffusion down gradient. Move through protein complex. Na+ pump expend energy. Antiport pump Na+ out, allow uptake of calcium.

23