Pulse Radiolysis of Lactate Dehydrogenase

Publisher Website
Google Scholar
Abstract
Pulse radiolysis has been used to investigate the rates and transient spectra for the reactions of free radicals with beef heart lactate dehydrogenase at pH 7. Analysis of the results leads to second-order rate-constants for eaq−, ·OH, ·I, ·Br2−, ·I2− and ·(CNS)2− which are, respectively, 24, 21, 10, 0·55, 0·43 and 0·15 in units of 1010 M−1 s−1 with uncertainties of ± 20 per cent. Those for ·I and ·I2− are similar to the corresponding rate-constants for the related enzyme alcohol dehydrogenase. The spectra of the transient species produced by ·OH, ·Br2− and ·(CNS)2− all showed evidence for reactions with tyrosine and tryptophan residues, and in general terms the magnitudes of the rate-constants appeared to increase with the oxidizing abilities of the radicals. The implication of the results for understanding the mechanism of deactivation by free radicals is discussed.