Effects of electron mediator charge properties on the reaction kinetics of hydrogenase from Chlamydomonas.
Anions modulate hydrogenase activity in cell-free preparations of Chlamydomonas reinhardtii, and this modulation is greatly influenced by the charge properties of the redox agent included to mediate electron transfer to hydrogenase. With cationic methyl viologen as the electron mediator, anions stimulate the maximum velocity of H2 production (e.g., a 320% increase in the presence of 1 M NaCl) but have little effect on the Km for methyl viologen. Conversely, when hydrogenase activity is mediated by polyanionic metatungstate or ferredoxin, H2 production is strongly inhibited by anions (e.g., 70-77% inhibition by 0.2 M NaCl). This inhibition is primarily due to a reduced affinity of hydrogenase for these mediators (as evidenced by a large increase in Km values), rather than a change in the maximum velocity of the reaction. Anions have little effect on the kinetics of hydrogenase activity mediated by zwitterionic sulfonatopropyl viologen, a redox agent with a nearly neutral net charge. These results suggest the presence of a cationic region near the active site of hydrogenase. This cationic region, probably due to lysine and/or arginine residues, may serve in vivo to facilitate the interaction between hydrogenase and ferredoxin, the polyanionic, physiological electron mediator.
D000838:Anions / Q000494:pharmacology; D001665:Binding Sites; D002696:Chlamydomonas / Q000201:enzymology*; D004563:Electrochemistry; D004579:Electron Transport / Q000187:drug effects*; D004789:Enzyme Activation / Q000187:drug effects; D005288:Ferredoxins / Q000494:pharmacology; D006864:Hydrogenase; D007700:Kinetics; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010269:Paraquat / Q000031:analogs & derivatives / Q000494:pharmacology; D014414:Tungsten / Q000494:pharmacology; D017973:Tungsten Compounds
D000838:Anions; D005288:Ferredoxins; C040584:N,N'-bis(sulfonatopropyl)-4,4'-bipyridyl; D017973:Tungsten Compounds; C040585:sodium metatungstate; D010088:Oxidoreductases; D006864:Hydrogenase; D010269:Paraquat; D014414:Tungsten