[Primary structure of 20S,22R-cholesterol-hydroxylating cytochrome P-450 from bovine adrenal cortex mitochondria. IV. Structure of peptides of thermolytic and limited tryptic hydrolysis of the fragment F1; peptides of cyanogen bromide hydrolysis of cytochrome P-450. Complete amino acid sequence].
A thermolytic hydrolysis of maleinated fragment F1 has been performed, resulted in isolation of 44 peptides; their complete amino acid sequence has been determined. Non-overlapping thermolytic peptides of fragment F1 involve 178 amino acid residues, which comprises about 71% of its amino acid sequence. Also, the cleavage and structural investigation of some tryptophan-containing peptides obtained from the limited trypsinolysis of fragment F1 were carried out; reconstitution of the polypeptide chain of the fragment is completed. The cyanogen bromide cleavage of carboxymethylated cytochrome P-450 was achieved and 17 peptides, comprising almost the whole polypeptide chain of the protein molecule (91%), was isolated. To investigate structure of the cyanogen bromide peptides, we hydrolysed them at tryptophan residues with trypsin, chymotrypsin, proteinase from Staphylococcus aureus, and BNPS-skatole. The data obtained and those published earlier led to the complete primary structure of the cholesterol-hydroxylating cytochrome P-450. The proteins polypeptide chain consists of 481 amino acid residues and has the precise molecular mass 56 407.7.
D000302:Adrenal Cortex / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000032:analysis; D002918:Chymotrypsin; D003488:Cyanogen Bromide; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D010450:Endopeptidases; D006868:Hydrolysis; D008666:Metalloendopeptidases; D008928:Mitochondria / Q000201:enzymology*; D010446:Peptide Fragments / Q000032:analysis*; D013250:Steroid Hydroxylases / Q000032:analysis*
D010446:Peptide Fragments; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D002786:Cholesterol Side-Chain Cleavage Enzyme; D010450:Endopeptidases; D002918:Chymotrypsin; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D003488:Cyanogen Bromide
Pervichnaia struktura 20S,22R-kholesteringidroksiliruiushchego tsitokhroma P-450 iz mitokhondriĭ kory nadpochechnikov byka. IV. Struktura peptidov termoliticheskogo i ogranichennogo tripticheskogo gidrolizov fragmenta F1; peptidy bromtsianovogo rasshchepleniia tsitokhroma P-450. Polnaia aminokislotnaia posledovatel'nost'.