title: Inactivation of Hydrogenase in Cell-free Extracts and Whole Cells of Chlamydomonas reinhardi by Oxygen.
abstract: O(2) irreversibly inactivates hydrogenase from Chlamydomonas reinhardi. The mechanism for the inactivation involves the reaction of one molecule of hydrogenase with one molecule of O(2) (or two oxygen atoms) in the transition complex of the rate-limiting step. The second order rate constant for this reaction is 190 atmospheres(-1) minute(-1) (1.4 x 10(5) molar(-1) minute(-1)). At levels above 0.01 atmosphere O(2), the increased numbers of O(2) molecules may compete for the site of inactivation hindering the proper orientation for inactivation of any one O(2) molecule and resulting in lowered rates of inactivation.CO is a reversible inhibitor of hydrogenase acting competitively against H(2). The K(i) for CO is 0.0010 atmosphere. CO antagonizes O(2) inactivation. In a period when complete inactivation by O(2) would usually occur, the presence of CO greatly reduces the inactivation rate.After 3 hours of adaptation in whole cells, the presence of H(2) lowers the rate of deadaptation of hydrogenase. Inasmuch as H(2) promotes increased O(2) uptake the cellular concentration of O(2) is likely to be lower. After 48 hours of adaptation O(2) uptake is reduced even when H(2) is present and the pattern of deadaptation under O(2) with and without H(2) and CO is qualitatively the same as observed for the inactivation of cell-free hydrogenase. The mechanism of inactivation of cell-free hydrogenase by O(2) may be the same as the mechanism for loss of hydrogenase during deadaptation in whole algal cells.
authors: Institute for Photobiology of Cells and Organelles, Brandeis University, Waltham, Massachusetts 02154.
authors: D L
authors: DL
authors: Erbes
authors: D
authors: D
authors: King
authors: M
authors: M
authors: Gibbs