title: Proteolysis of L-(+)-lactate cytochrome c oxidoreductase (cytochrome b2) extracted from Saccharomyces cerevisiae and Hansenula anomala yeasts.
abstract: The L-(+)-Lactate:cytochrome c oxidoreductase or cytochrome b2 from the yeasts Saccharomyces cerevisiae and Hansenula anomala were partially hydrolysed in various concentrations of trypsin. Conditions were found which allowed the isolation from the Hansenula enzyme of a 140 000 +/- 10 000-dalton flavoprotein. The prosthetic flavin groups were still reducible by substrate (spectroscopic evidence) but the flavoprotein was unable to form a complex with cytochrome c, the physiological acceptor in the enzymatic reaction. No such flavoprotein units could be found during proteolysis of the Saccharomyces enzyme. The heme prosthetic group of the Hansenula enzyme remained bound to a 15 500 +/- 1000-dalton protein unit which was larger than, but very similar to, the well known 'cytochrome b2 core' of the Saccharomyces enzyme. Moreover, the degradation of different enzyme samples by contaminated proteases allowed the isolation of a particular form of Hansenula enzyme: each tetramer had, on the mean, four bound flavins and only two heme groups. These molecules completely retained their ability to form a complex with cytochrome c.
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D003574:Cytochrome c Group; D003580:Cytochromes / Q000302:isolation & purification / Q000378:metabolism; D005415:Flavins / Q000032:analysis; D006418:Heme / Q000032:analysis; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010843:Pichia / Q000201:enzymology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013045:Species Specificity; D014357:Trypsin
chemical_list: D003574:Cytochrome c Group; D003580:Cytochromes; D005415:Flavins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D006418:Heme; D007770:L-Lactate Dehydrogenase; D014357:Trypsin