Characterization of a fourth tungsten-containing enzyme from the hyperthermophilic archaeon Pyrococcus furiosus.
Pyrococcus furiosus grows optimally near 100 degrees C using peptides and carbohydrates as carbon sources, and it reduces elemental sulfur (S(0)), if present, to H(2)S. Tungsten (W), an element rarely used in biology, is required for optimal growth, and three different tungsten-containing enzymes have been previously purified from this organism. They all oxidize aldehydes of various types and are thought to play primary roles in the catabolism of sugars or amino acids. Here, the purification of a fourth tungsten-containing enzyme, termed WOR 4, from cell extracts of P. furiosus grown with S(0) is described. This was achieved by monitoring through multiple chromatography steps the W that is not associated with the three characterized tungstoenzymes. The N-terminal sequence of WOR 4 and the approximate molecular weight of its subunit determined electrophoretically (69,000) correspond to the product of an ORF (PF1961, wor4) present in the complete genome sequence of P. furiosus. WOR 4 is a homodimer and contains approximately one W, three Fe, three or four acid-labile sulfide, and one Ca atom per subunit. The visible and electron paramagnetic resonance spectra of the oxidized and reduced enzyme indicate the presence of an unusual iron-sulfur chromophore. WOR 4 does not oxidize aliphatic or aromatic aldehydes or hydroxy acids, nor does it reduce keto acids. Consistent with prior microarray data, the protein could not be purified from P. furiosus cells grown in the absence of S(0), suggesting that it may have a role in S(0) metabolism.
Roy
Roopali
R
Department of Biochemistry and Molecular Biology and Center for Metalloenzyme Studies, University of Georgia, Athens 30602, USA.
Adams
Michael W W
MW
D000445:Aldehyde Oxidoreductases / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D019843:Archaeal Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D002384:Catalysis; D008970:Molecular Weight; D020027:Pyrococcus furiosus / Q000201:enzymology*; D013455:Sulfur / Q000378:metabolism; D014414:Tungsten / Q000032:analysis*
D019843:Archaeal Proteins; D013455:Sulfur; D000445:Aldehyde Oxidoreductases; C080010:aldehyde ferredoxin oxidoreductase; D014414:Tungsten