Molybdenum and vanadium do not replace tungsten in the catalytically active forms of the three tungstoenzymes in the hyperthermophilic archaeon Pyrococcus furiosus.
Three different types of tungsten-containing enzyme have been previously purified from Pyrococcus furiosus (optimum growth temperature, 100 degrees C): aldehyde ferredoxin oxidoreductase (AOR), formaldehyde ferredoxin oxidoreductase (FOR), and glyceraldehyde-3-phosphate oxidoreductase (GAPOR). In this study, the organism was grown in media containing added molybdenum (but not tungsten or vanadium) or added vanadium (but not molybdenum or tungsten). In both cell types, there were no dramatic changes compared with cells grown with tungsten, in the specific activities of hydrogenase, ferredoxin:NADP oxidoreductase, or the 2-keto acid ferredoxin oxidoreductases specific for pyruvate, indolepyruvate, 2-ketoglutarate, and 2-ketoisovalerate. Compared with tungsten-grown cells, the specific activities of AOR, FOR, and GAPOR were 40, 74, and 1%, respectively, in molybdenum-grown cells, and 7, 0, and 0%, respectively, in vanadium-grown cells. AOR purified from vanadium-grown cells lacked detectable vanadium, and its tungsten content and specific activity were both ca. 10% of the values for AOR purified from tungsten-grown cells. AOR and FOR purified from molybdenum-grown cells contained no detectable molybdenum, and their tungsten contents and specific activities were > 70% of the values for the enzymes purified from tungsten-grown cells. These results indicate that P. furiosus uses exclusively tungsten to synthesize the catalytically active forms of AOR, FOR, and GAPOR, and active molybdenum- or vanadium-containing isoenzymes are not expressed when the cells are grown in the presence of these other metals.
Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602, USA.
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D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001105:Archaea / Q000201:enzymology*; D004578:Electron Spin Resonance Spectroscopy; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D008982:Molybdenum / Q000032:analysis*; D010088:Oxidoreductases / Q000378:metabolism; D014414:Tungsten / Q000032:analysis* / Q000378:metabolism; D014639:Vanadium / Q000032:analysis*
D014639:Vanadium; D008982:Molybdenum; D010088:Oxidoreductases; D000445:Aldehyde Oxidoreductases; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; C080010:aldehyde ferredoxin oxidoreductase; C080011:formaldehyde ferredoxin oxidoreductase; C093132:glyceraldehyde-3-phosphate ferredoxin oxidoreductase; D014414:Tungsten