Presentation on theme: "Membrane structure Membrane Transport of Small Molecules"— Presentation transcript:
1 Membrane structureMembrane Transport of Small MoleculesElectrical Properties of Membranes
2 Importance of cell membranes Plasma membrane separates cellular components from the environment.Allows organelles to execute specialized functions by keeping the contents of the organelle separate from the rest of the cell.Provides boundaries that establish electrochemical gradients. These are used to make ATP and to generate nerve impulses.
3 Biological membranes are thin films composed mainly of amphipathic lipids and proteins. Most of the molecules are held together by non-covalent interactions.
4 phosphatidylcholin Polar head (phospholipid) Hydrocarbon tail derived from fatty acid.Nonpolar tailUnsaturated hydrocarbon tailsaturated hydrocarbon tail
7 Amphipathic molecules pack so as to minimize the interaction between water and the nonpolar parts. The two hydrocarbon tails give phospholipids a cylindrical shape that causes the molecules to pack as a bilayer in water.
9 CholesterolAnother type of lipid called cholesterol decreases fluidity because it restricts the movement of the hydrocarbon chains.Important chemical characteristics of cholesterol:Hydroxyl group constitutes the polar head group.OH is attached to rigid steroid ring.One hydrocarbon tail.
28 Transmembrane alpha helix The hydrophobic core of the bilayer is 3 nm thick.One turn of the helix spans 0.54 nm and there are 3.5 a.a./turn. Hence, an transmembrane domain composed composed of an alpha-helix is around 20 amino acids long.
31 GlycophorinAdditional characteristics of many alpha helical transmembrane proteins:SH groups facing the cytoplasm are reduced and those facing outside the cell are oxidized to S-S.Sugars are covalently attached to certain amino acid side chains in regions that face outside the cell.
32 *Protein 3 is known as a beta barrel and is composed of a beta sheet curved into a barrelMuch rarer than the alpha helix transmembrane domain - limited to the outer membranes of bacteria, chloroplasts, and mitochondria.One well-known representative is a protein called Porin. The inside of the barrel is lined with polar amino acid side chains and the outside of the barrel is lined with nonpolar amino acid side chains.The polar groups in the peptide backbone are “hidden” by hydrogen bonding with antiparallel strands.
35 DetergentsMild nonionic detergent that dissolves membranes without unfolding proteins.Both are amphipathicIonic detergent that dissolves membranes and unfolds proteins.
36 SDS-PAGE (Polyacrylamide Gel Electrophoresis) After electrophoretic separation, proteins in thegel can be stained with a dye (Coomassie blue)which binds to protein.- Binds to most proteins (hydrophobic interactions?) inamounts roughly proportional to the molecular weight ofthe protein (~ 1 SDS molecule for every 2 aa residues).- Bound SDS contributes large -ve charge >> intrinsic chargeof protein, so proteins are separated almost exclusivelyon the basis of molecular weight.
37 DetergentsA major advance in the study of membrane proteins involved in transport was the discovery that the proteins would spontaneously assemble with purified lipids in vesicles sometimes called “proteoliposomes”. “Liposome” indicates a vesicle made of lipids and “proteo” indicates that proteins are present.
38 Membrane proteins form complexes eg. “photosynthetic reaction center”(from the bacterium Rhodopseudomonas)- has 4 subunits, 3 of which containa-helical segments that span themembrane (total: 11)- helical segments are rich innon-polar aa, whose hydrophobicside chains interact with themembrane lipids that surroundthe complex[In red: non-protein prosthetic groups and electon-carrying isoprenoid compounds called quinones]
39 Fluorescence recovery after photobleaching provides evidence that many proteins undergo lateral diffusion.
47 Transport of molecules that are impermeable to the lipid bilayer is achieved by two main classes of membrane transport proteins.Conformational change “carries” the solute across the membrane.Aqueous pore provides passage way so solute can diffuse through the membrane.
48 Four ways of molecules and ions across the membrane
50 Carrier ProteinsA carrier protein binds solute on one side of a membrane, undergoes a conformational change, and releases the solute on the other side of the membrane.
51 A carrier protein resembles an enzyme Enzyme Carrier proteinBinding sites for substrates Binding sites for soluteChemically transforms substrate Transports soluteReaction rate saturate at high Transport rate saturates at highsubstrate concentration solute concentration
52 By coupling the conformational change to a source of energy, a carrier protein can perform active transport.
53 Depending on how many solute molecules are transported and in what direction, carrier proteins are dubbed uniporters, symporters, or antiporters.
54 Na+-K+ pump (aka Na+-K+ ATPase) in the plasma membrane is an antiporter that performs active transport.This protein establishes a concentration gradient for Na+ that is low inside the cell and high outside, and a concentration gradient for K+ this high inside the cell and low outside.
55 Direct active transport: The Na+-K+ pump (a P-class pump)
56 The Na+ gradient generated by the Na+ - K+ ATPase powers the transport of glucose into the cell by a Na+ -driven glucose symporter.The energetically favorable movement of Na+ down its electrochemical gradient is coupled to the energetically unfavorable transport of glucose up its concentration gradient. Hence, glucose is being subjected to active transport.
57 Glucose transport: analogy with enzyme Sout + Tk1k-1Sout.Tk2k-2Sin.Tk3k-3Sin + TSUBSTRATEPRODUCT“ENZYME”Can be described by:Kinetics: rate equations give an expression analogous to Michaelis-Menten equation:V0 =Vmax [S]outKt + [S]outV0 = accumulation of glucose inside cell when its concentration outside is [S]outKt = measure of affinity of transporter for S
58 Three carrier proteins, appropriately positioned in the plasma membrane, function to transport glucose across the intestinal epithelium.
63 Ion channels are ion-selective because of narrow region called the “selectivity filter”. side viewRRtop view - each circle represents an end-on view of an alpha-helix and the R-groups radiating outwards are amino acid side chains.RRRRRRRRRRRRR
65 Stryer Fig. 13.27 Selectivity of a sodium channel.
66 How can a K+ channel discriminate between K+ and Na+ since they both have the same charge and Na+ is smaller than K+?The answer: as the ion passes through the selectivity filter, the ion must shed water. Carbonyl oxygens with partial negative charge can take the place of water for K+ but Na+ is too small. Hence, Na+ favors remaining associated with water and hydrated ion is too large to fit through the selectivity filter.
67 Ion channels fluctuate between open and closed state to regulate ion flow. Voltage-gated channels respond to the membrane potential.Ligand-gated channels respond to association of small molecules called ligands.Mechanically gated channels respond to movement.
68 Establishing a resting membrane potential in animal cells results from the coordinated action of carrier proteins and ion channels.1. Na+-K+ ATPase concentrates K+ inside the cell and Na+ outside (active transport).2. K+ leak channels allow K+ to diffuse out of the cells, down the concentration gradient (passive transport).3. Negative charge left behind in the cytoplasm counteracts the efflux of K+ so only a very small amount (1/100,000) K+ leak out.4. The efflux of the K+ is sufficient to generate a membrane potential of approximately -100 mV - positive outside and negative inside.The resting potential is defined as the membrane potential occurring when there is no net flow of ions. Cells typically have resting potentials between -20 and -200 mV. The major contributor is the K+ leak channel, but other channels account for the range of potentials observed.
69 Resting Membrane Potential: The Nernst equation
70 Nerve cell (Neuron)Dendrites - cell protrusions that receive signals from axons.Cell body - location of nucleus.Axon - single long protrusion that sends signal away from the cell body.A nerve impulse results from electrical disturbances in the plasma membrane that spread from one part of the cell to another. The electrical disturbance is called an action potential and it consists of a wave of membrane depolarization that moves down the axon.
71 Membrane depolarizations spread passively only short distances
72 Voltage-gated channels are the key to an action potential. Depolarization to a specific membrane potential causes the channels to open. This specific membrane potential can be thought of as the threshold that must be reached to get the channel to open.
73 The structure and function of the voltage-gated Na+ channel Figure 21-13
74 A threshold depolarization initiates the action potential. When a nerve receives a signal, a modest depolarization of the membrane occurs. If this depolarization reaches the threshold, all the voltage-gated Na+ channels experiencing this threshold depolarization will open simultaneously. Na+ rushes into the cell causing a rapid and large depolarization of the membrane. This rapid and large depolarization is the action potential.
75 The channel cycle of closed, opened, inactivated along the axon results in propagation of the action potential.
76 Initiation of an action potential requires that something cause the membrane to depolarize to the threshold potential. The process begins at the synapse.presynaptic cellpostsynaptic cell
77 Neurotransmitters are small molecules that transmit impulses at chemical synapses
79 A single nerve cell combines the excitatory and inhibitory signals received from numerous other neurons to control the frequency with which action potentials are generated.The presynaptic terminals are derived from numerous other neurons. Ligand-gated ion channels are concentrated at the synapses on the postsynaptic cell (colored yellowish-brown).Neurotransmitters released into each synapse will generate a postsynaptic potential (PSP).The voltage-gated Na+ channels are concentrated at the axon hillock.An action potential is initiated at the axon hillock when the sum of excitatory and inhibitory PSP depolarize the axon hillock to the threshold potential.
80 The greater the “combined PSP”, the more frequent action potentials are generated at the axon hillock. Note that the magnitude of the change in membrane potential during an action potential is constant because the membrane potential is limited by the Na+ concentration.
81 Myelination increases the speed and efficiency of action potential propagation. Glial cells (diagrammed in red) surrounding the axon produce an electrical insulation rich in glycolipids.Nodes of Ranvier located at regular intervals are openings in the insulation where Na+ channels are concentrated.Influx of Na+ at one node results in depolarization at the next node due to the rapid diffusion of Na+ in the cytoplasm. This triggers an action potential that leads to rapiddepolarization of the next node.The action potential jumps from node to node by a process called saltatory conduction.
82 Action potentials travel rapidly from one node to the next Figure 21-18
83 The neuromuscular junction is an example of how the action potential from the nerve triggers a response in another cell.
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