Selenophosphate synthetase: enzyme labeling studies with [gamma-32P]ATP, [beta-32P]ATP, [8-14C]ATP, and [75Se]selenide

Archives of Biochemistry and Biophysics
S Y Liu, T C Stadtman

Abstract

Selenophosphate synthetase catalyzes a reaction in which ATP and selenide are converted to H3SeP03, H3P04, and AMP in a 1:1:1 ratio. Selenophosphate is derived from the gamma phosphoryl group and orthophosphate from the beta phosphoryl group of ATP. In the absence of selenide, a slow reaction in which ATP is converted quantitatively to 2 H3P04 and AMP occurs. Labeling experiments carried out to detect a putative enzyme-bound pyrophosphate intermediate in the overall reaction showed that up to 0.6 equivalent of the 32P label from [gamma-32P]ATP was bound to protein under enzyme turnover conditions, but only a negligible amount of 32P from [beta-32P]ATP was present. Thus, no Enz-PP intermediate was present in a detectable amount under the experimental conditions used. Isolated enzyme samples contained 75Se from 75Se-labeled selenide and [14C]AMP from [8-14C]ATP in amounts similar to the bound 32P from [gamma-32P]ATP, suggesting that two of the final products, selenophosphate and AMP, were the radioactive compounds detected in these experiments.

References

Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·W LeinfelderA Böck
Jan 1, 1995·Methods in Enzymology·R S Glass, T C Stadtman
Jul 19, 1994·Proceedings of the National Academy of Sciences of the United States of America·I Y Kim, T C Stadtman

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Citations

Jun 10, 2008·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Eiko MatsumotoShigeyuki Yokoyama
Apr 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·H WalkerT C Stadtman
Dec 28, 1999·BioFactors·G M Lacourciere
Sep 9, 2008·Journal of Molecular Biology·Yuzuru ItohShigeyuki Yokoyama
Dec 1, 2004·EcoSal Plus·Matt D Wolfe

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