title: Selenophosphate synthetase: detection in extracts of rat tissues by immunoblot assay and partial purification of the enzyme from the archaean Methanococcus vannielii.
abstract: In Escherichia coli and Salmonella typhimurium it has been shown that selenophosphate serves as the selenium donor for the conversion of seryl-tRNA to selenocysteyl-tRNA and for the synthesis of 2-selenouridine, a modified nucleoside present in tRNAs. Although selenocysteyl-tRNA also is formed in eukaryotes and is used for the specific insertion of selenocysteine into proteins, the precise mechanism of its biosynthesis from seryl-tRNA in these systems is not known. Because selenophosphate is extremely oxygen labile and difficult to identify in biological systems, we used an immunological approach to detect the possible presence of selenophosphate synthetase in mammalian tissues. With antibodies elicited to E. coli selenophosphate synthetase the enzyme was detected in extracts of rat brain, liver, kidney, and lung by immunoblotting. Especially high levels were detected in Methanococcus vannielii, a member of the domain Archaea, and the enzyme was partially purified from this source. It seems likely that the use of selenophosphate as a selenium donor is widespread in biological systems.
authors.affiliation: Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-0320, USA.
authors.forename: I Y
authors.initials: IY
authors.lastname: Kim
authors.forename: T C
authors.initials: TC
authors.lastname: Stadtman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification* / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D029721:Drosophila Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D015151:Immunoblotting / Q000379:methods; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D017017:Methanococcus / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010770:Phosphotransferases; D051381:Rats; D012486:Salmonella typhimurium / Q000201:enzymology; D017386:Sequence Homology, Amino Acid
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D029721:Drosophila Proteins; D010770:Phosphotransferases; C062762:selenophosphate synthetase