Abstract
Auto‐ADP‐ribosylation of the glycolytic enzyme glyceraldehyde‐3‐phosphate dehydrogenase (GraPDH) has recently been demonstrated to be dramatically stimulated in the presence of nitric oxide. In order to obtain insight into the sequence of events leading to ADP‐ribosylation of GraPDH, we studied the target amino acid, the nucleotide cofactor requirement, pH dependency and the stoichiometry of the reaction. Basal as well as stimulated ADP‐ribose transfer is inhibited by the SH‐group alkylating reagent, N‐ethylmaleimide. Furthermore, the radiolabel of auto‐[32P]ADP‐ribosylated GraPDH is removed by treatment with HgCl2, suggesting an ADP‐ribose–cysteine bond. Several indirect and direct mechanistic considerations point to NAD+ as the only cofactor for the ADP‐ribosylation reaction, excluding the possibility of a reaction sequence involving a NAD‐glycohydrolase(s) followed by nonenzymatic ADP‐ribose transfer to GraPDH. Optimal ADP‐ribosylations were carried out at alkaline pH values using 10 μM free NAD+ as the sole nucleotide cofactor. Bovine serum albumin with an S‐nitrosylated SH group can serve as a model of ADP‐ribose transfer from NAD+ and suggests that the nitric‐oxide‐modified SH group (S‐nitrosylated SH group) is a prerequisite for the reaction.
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Dates
Type | When |
---|---|
Created | 20 years, 5 months ago (March 4, 2005, 2:36 a.m.) |
Deposited | 1 year, 9 months ago (Oct. 30, 2023, 11:38 p.m.) |
Indexed | 1 year, 1 month ago (July 6, 2024, 10:34 a.m.) |
Issued | 32 years, 9 months ago (Nov. 1, 1992) |
Published | 32 years, 9 months ago (Nov. 1, 1992) |
Published Online | 20 years, 5 months ago (March 3, 2005) |
Published Print | 32 years, 9 months ago (Nov. 1, 1992) |
@article{DIMMELER_1992, title={Characterization of a nitric‐oxide‐catalysed ADP‐ribosylation of glyceraldehyde‐3‐phosphate dehydrogenase}, volume={210}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1992.tb17422.x}, DOI={10.1111/j.1432-1033.1992.tb17422.x}, number={1}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={DIMMELER, Stefanie and BRÜNE, Bernhard}, year={1992}, month=nov, pages={305–310} }