Cell-free synthesis of a flavoprotein containing the 8 alpha-(N3-histidyl)-riboflavin linkage.
6-Hydroxy-D-nicotine oxidase is an inducible flavorprotein of Arthrobacter oxidans in which one mole of FAD is bound covalently to the polypeptide chain. During cell-free translation of polysomes from nicotine-induced A. oxidans cells in the presence of an Escherichia coli (MRE 600) supernatant fraction, labelled FAD, leucine and histidine were incorporated into 6-hydroxy-D-nicotine oxidase in the same ratio found in the enzyme isolated from whole cells. This indicates that one mole FAD is covalently attached per mol of 6-hydroxy-D-nicotine oxidase synthesized in vitro. In the native enzyme the coenzyme molecule is bound via its 8 alpha-methyl group to the N-3 atom of a histidyl residue. From the translation products an aminoacyl-riboflavin was isolated and its identity with synthetic 8 alpha-(N3-histidyl)-riboflavin was shown.
D001173:Arthrobacter / Q000201:enzymology*; D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D005420:Flavoproteins / Q000096:biosynthesis*; D006639:Histidine / Q000032:analysis*; D007700:Kinetics; D008970:Molecular Weight; D010088:Oxidoreductases / Q000096:biosynthesis*; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D012256:Riboflavin / Q000378:metabolism*
D005420:Flavoproteins; D006639:Histidine; D010088:Oxidoreductases; C021413:6-hydroxy-D-nicotine oxidase; D012256:Riboflavin