title: Biosynthesis of selenophosphate.
abstract: Selenophosphate synthetase, the product of the selD gene, produces the highly active selenium donor, monoselenophosphate, from selenide and ATP. Positional isotope exchange experiments have shown hydrolysis of ATP occurs by way of a phosphoryl-enzyme intermediate. Although, mutagenesis studies have demonstrated Cys17 in the Escherichia coli enzyme is essential for catalytic activity the nucleophile in catalysis has not been identified. Recently, selenophosphate synthetase enzymes have been identified from other organisms. The human enzyme which contains a threonine residue corresponding to Cys17 in the E. coli enzyme, has been overexpressed in E. coli. The purified enzyme shows no detectable activity in the in vitro selenophosphate synthetase assay. In contrast, when the human enzyme is expressed to complement a selD mutation in E. coli, in the presence of 75Se, incorporation of 75Se into bacterial selenoproteins is observed. The inactive purified human enzyme together with the very low determined specific activity of the E. coli enzyme (83 nmol/min/mg) suggest an essential component for the formation of selenophosphate has not been identified.
authors.affiliation: Laboratory of Biochemistry, National Heart, Lung, Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: G M
authors.initials: GM
authors.lastname: Lacourciere
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000378:metabolism*; D029721:Drosophila Proteins; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D051379:Mice; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010710:Phosphates / Q000378:metabolism*; D010770:Phosphotransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018036:Selenium Compounds / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D029721:Drosophila Proteins; D010710:Phosphates; D018036:Selenium Compounds; C076818:selenophosphate; D010770:Phosphotransferases; C062762:selenophosphate synthetase