Pump driven by ATP hydrolysismechanism (see also Fig. 12-34 in Nelson & Cox) involves reversible phosphorylation of an Asp residue on the enzyme, and 2 conformations of the enzyme:Conformation I: high affinity for Na+, low affinity for K+, "open" to inside of cell.Conformation II: low affinity for Na+, high affinity for K+, "open" to outside of cell.Transfer of phosphate group from ATP to enzyme (releasing ADP as a product) triggers conformational change in enzyme -- phosphorylated enzyme predominantly in conformation II.Hydrolysis of phosphate group from the enzyme triggers return to original conformation (I).Mechanism of the Na+-K+ pump, starting on upper left (like Nelson & Cox Fig. 12-34)Unphosphorylated enzyme (EnzI) binds 3 Na+ from inside cell[EnzI •3Na+] is phosphorylated (on Asp residue), generating second conformation.EnzII-P releases 3 Na+ ions outside and binds 2 K+ ions from outside cell.[EnzII-P •2K+] has phosphate hydrolyzed off (inside cell).Unphosphorylated enzyme switches to conformation I (EnzI), releasing 2K+ inside cell, now ready to bind 3 Na+ again.This animation shows operation of the pump. Upper side = outside of cell; lower side = cytosol. The colored ball represents ATP; the three yellow diamonds Na+ and the two red diamonds K+.Ion gradients (and electrical potentials/gradients) provide the energy for SECONDARY ACTIVE TRANSPORT.(Ionophores dissipate ion gradients and thus are poisons, or antibiotics for microorganisms.)COTRANSPORT processes that utilize a favorable gradient for one compound to drive the uptake of a second compound.Sodium-glucose cotransport across apical surface of intestinal epithelial cells is one example, accumulating Glc in cell against its concentration gradientFig. 12-36 (Nelson & Cox, Lehninger Principles of Biochemistry, 3rd ed., 2000): Glucose transport in intestinal epithelial cellsGlucose import from intestine made possible by Na+-K+ ATPase (shown on right side of cell), which generates/maintains both high Na+ concentration outside cell and charge gradient (electrical potential) that both favor Na+ import through Na+-glucose symporter.Permits epithelial cells to concentrate glucose from intestine to 30,000x the intestinal concentrationResulting high concentration of glucose within cell passes "down" its concentration gradient through basal surface of cell into blood via GluT2 transporter (facilitated diffusion, uniport system).Many other secondary active transport systems are known, especially in mitochondrial membrane.Other examples of secondary active transport:Sodium-calcium exchanger of animal cell membranes: antiporter that couples downhill flow of 3 Na+ into cell with uphill extrusion of 1 Ca2+ out of the cell (Na+ gradient was generated by the Na+-K+ ATPase.)Lactose permease of E. coli: symporter that uses H+ gradient across E. coli membrane (generated by fuel oxidation and electron transport) to let protons flow down their concentration gradient back into the cell, bringing lactose into the cell against a concentration gradient (see Fig. 12-35 in Nelson & Cox, Lehninger Principles). This perspective/commentary from Science 301, 603-4 (1 Aug. '03) (PDF) discusses the crystal structure and the function of lactose permease (lacY), with references.
đang được dịch, vui lòng đợi..
