Controlled digestion with trypsin as a structural probe for the N-terminal peptide of soluble and membranous cytochrome c.
When purified bovine cytochrome c1 is digested with trypsin under controlled conditions, the heme polypeptide is preferentially converted from a species of molecular weight 30,600 to a heme polypeptide of molecular weight 29,000. The trypsin sensitive peptide bond is located in the N-terminal region of the cytochrome. Both the reduced and oxidized cytochrome are susceptible to hydrolysis by trypsin at the same locus, but the reduced cytochrome is cleaved at an initial rate approximately twofold greater than the oxidized cytochrome. Membranous cytochrome c1, as occurring in cytochrome b-c1 complex or succinate-cytochrome c reductase complex, is not susceptible to trypsin proteolysis under similar conditions, nor after more extensive treatment of the membranes with trypsin, in spite of the fact that cytochrome c1 presumably comes into contact with cytochrome c at the membrane surface during electron transport. These findings are consistent with a model for the structure of cytochrome c1 in situ in which the cytochrome is an integral membrane protein, located primarily in the membrane continuum, while still having the heme-containing portion of the protein available at the membrane surface for electron transfer to cytochrome c.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D003579:Cytochrome Reductases; D003574:Cytochrome c Group; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000032:analysis; D007700:Kinetics; D008566:Membranes / Q000201:enzymology*; D008931:Mitochondria, Muscle / Q000201:enzymology*; D009097:Multienzyme Complexes; D009206:Myocardium; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000032:analysis; D014357:Trypsin
D000596:Amino Acids; D003574:Cytochrome c Group; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D006418:Heme; D003579:Cytochrome Reductases; D014357:Trypsin