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A new mechanism of acyl group transfer in the reaction catalyzed by D-glyceraldehyde-3-phosphate dehydrogenase

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Abstract

D-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) catalyzes the oxidative phosphorylation of its substrate in a two-step reaction. Asa result of the first, oxidativestep, the covalent intermediate where in 3-phosphoglyceroyl moiety is bound to Cys149 of the active center is subjected to nucleophilic attack by inorganic phosphate, but remains resistant to hydrolytic decomposition. This ensures tight coupling of oxidation with phosphorylation in glycolysis. In this article, we present the experimental evidence for the conversion of GAPDH into a form capable of performing the reaction in the absence of inorganic phosphate. The structural basis for this conversion is the oxidation of a cysteine residue (probably Cys 153) into a sulfenic acid derivative under mild conditions to affect the integrity of the essential Cys 149. As a result, an intram olecular transfer of 3-phosphoglyceroyl group from the active center Cys 149 to Cys 153 becomes possible with subsequent hydrolysis of the sulfenyl carboxylate intermediate.

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Authors and Affiliations

  1. A. N. Belozersky Institute of Physico-Chemical Boilogy, Moscow State University, 119899, Moscow, Russia

    Elena V. Schmalhausen, Vladimir I. Muronetz & Natalya K. Nagradova

Authors
  1. Elena V. Schmalhausen
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  2. Vladimir I. Muronetz
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  3. Natalya K. Nagradova
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Correspondence to Vladimir I. Muronetz.

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Schmalhausen, E.V., Muronetz, V.I. & Nagradova, N.K. A new mechanism of acyl group transfer in the reaction catalyzed by D-glyceraldehyde-3-phosphate dehydrogenase. Appl Biochem Biotechnol 88, 211–220 (2000). https://doi.org/10.1385/ABAB:88:1-3:211

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  • DOI: https://doi.org/10.1385/ABAB:88:1-3:211

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