Structure of the flavin site of [formula omitted] flavocytochrome [formula omitted

Abstract

Two flavin peptides have been isolated from Chromatium cytochrome c 552 by digestion with pepsin and with trypsin-chymotrypsin, respectively. Acid hydrolysis and aminopeptidase digestion of the peptic peptide shows the presence of 1 mole each of threonine and cysteine and 2 of tyrosine, per mole of FAD. Edman degradation gives the sequence: tyr-thr-cys (flavin)-tyr. Tryptic-chymotryptic digestion yields a flavin tripeptide of the structure: thr-cys (flavin)-tyr. The N-terminal tyrosine in the peptic tetrapeptide shows a strong interaction with the flavin, as judged by CD spectra, and this may account for the resistance of the bridge sulfur to oxidation by cold performic acid. Both peptides show abnormally low fluorescence in the pure state (1 to 5% relative to riboflavin) and positive iodoplatinic test; the peptic peptide yields a positive, the trypticchymotryptic peptide negative ninhydrin reaction. The electrophoretic mobility of the major product of aminopeptidase digestion (presumably the thiazolidine form of cysteinyl-8α-FAD thiohemiacetal) and other properties of the two peptides are in accord with previous suggestions that the linkage of the flavin to the peptide is a thiohemiacetal.