To elucidate the catalytic mechanism of monodehydroascorbate (MDA) reductase from cucumber, its interaction with MDA radical was investigated by the use of pulse radiolysis. When approximately equimolar MDA radical to the fully reduced MDA reductase was generated, the fully reduced enzyme reacted first with MDA radical to form the red semiquinone, and the semiquinone further reacted with MDA radical to form the oxidized enzyme. At a low ratio (<20) of MDA radical to enzyme concentration, the fully reduced enzyme reacted quantitatively with MDA radical to form the semiquinone with a second-order rate constant of 2.6 10M s at pH 7.4. At excess MDA radical to enzyme concentration, a similar rate constant was obtained from the decay of MDA radical. These results suggest that the reaction of the semiquinone with MDA radical occurs at the same rate or rate-limiting step of the oxidation of the fully reduced enzyme by MDA radical. The rate constants decreased with an increase in NaCl concentration, suggesting that the localization of cationic groups of amino acid residue near the active site may provide electrostatic guidance to the anionic substrate of MDA radical.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				W.-C. Hou, Y.-T. Wang, Y.-H. Lin, L.-J. Hsiao, T.-E. Chen, C.-W. Wang, and H. Dai A complex containing both trypsin inhibitor and dehydroascorbate reductase activities isolated from mitochondria of etiolated mung bean (Vigna radiata L. (Wilczek) cv. Tainan No. 5) seedlings J. Exp. Bot., April 1, 2000; 51(345): 713 - 719. [Abstract] [Full Text] [PDF]

				K. Kobayashi, M. Tsubaki, and S. Tagawa Distinct Roles of Two Heme Centers for Transmembrane Electron Transfer in Cytochrome b561 from Bovine Adrenal Chromaffin Vesicles as Revealed by Pulse Radiolysis J. Biol. Chem., June 26, 1998; 273(26): 16038 - 16042. [Abstract] [Full Text] [PDF]