Molybdopterin in carbon monoxide oxidase from carboxydotrophic bacteria.
The carbon monoxide oxidases (COXs) purified from the carboxydotrophic bacteria Pseudomonas carboxydohydrogena and Pseudomonas carboxydoflava were found to be molybdenum hydroxylases, identical in cofactor composition and spectral properties to the recently characterized enzyme from Pseudomonas carboxydovorans (O. Meyer, J. Biol. Chem. 257:1333-1341, 1982). All three enzymes exhibited a cofactor composition of two flavin adenine dinucleotides, two molybdenums, eight irons and eight labile sulfides per dimeric molecule, typical for molybdenum-containing iron-sulfur flavoproteins. The millimolar extinction coefficient of the COXs at 450 nm was 72 (per two flavin adenine dinucleotides), a value similar to that of milk xanthine oxidase and chicken liver xanthine dehydrogenase at 450 nm. That molybdopterin, the novel prosthetic group of the molybdenum cofactor of a variety of molybdoenzymes (J. Johnson and K. V. Rajagopalan, Proc. Natl. Acad. Sci. U.S.A. 79:6856-6860, 1982) is also a constituent of COXs from carboxydotrophic bacteria is indicated by the formation of identical fluorescent cofactor derivatives, by complementation of the nitrate reductase activity in extracts of Neurospora crassa nit-l, and by the presence of organic phosphate additional to flavin adenine dinucleotides. Molybdopterin is tightly but noncovalently bound to the protein. COX, sulfite oxidase, xanthine oxidase, and xanthine dehydrogenase each contains 2 mol of molybdopterin per mol of enzyme. The presence of a trichloroacetic acid-releasable, so-far-unidentified, phosphorous-containing moiety in COX is suggested by the results of phosphate analysis.
D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002645:Chickens; D003067:Coenzymes; D005260:Female; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis; D008099:Liver / Q000201:enzymology; D008667:Metalloproteins; D008892:Milk / Q000201:enzymology; D008970:Molecular Weight; D008982:Molybdenum / Q000032:analysis*; D000090006:Molybdenum Cofactors; D010084:Oxidation-Reduction; D050862:Oxidoreductases Acting on Sulfur Group Donors; D011549:Pseudomonas / Q000201:enzymology*; D011621:Pteridines / Q000032:analysis*; D013045:Species Specificity; D013053:Spectrophotometry; D014968:Xanthine Dehydrogenase
D000445:Aldehyde Oxidoreductases; D003067:Coenzymes; D005182:Flavin-Adenine Dinucleotide; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D050862:Oxidoreductases Acting on Sulfur Group Donors; D011621:Pteridines; D014968:Xanthine Dehydrogenase; C022261:molybdopterin; C033598:carbon monoxide-methylene blue oxidoreductase