Isolation and characterization of the cyanogen bromide fragments of lobster arginine kinase (homarus vulgaris).
Arginine kinase was aminoethylated in order to block the five free thiol groups on the native enzyme, and then submitted to BrCN cleavage. The BrCN resulting peptides were soluble in propionic acid (10 percent) and subsequently submitted to gel-filtration. The large polypeptide subfractions were citraconylated and resubmitted to differnt gelchromatographies, whereas the short peptide subfractions were submitted to preparative paper electrochromatographies. Eight peptides of 2, 11, 17, 25, 61, 82, 86 and 132 amino acid residues were isolated, one of which is the overlapping of two peptides. The amino acid composition and the end group of all the isolated peptides were established. The short peptides (2, 11 and 17 residues) were sequenced. All peptides possess homoserine at C-terminal position because one methionyl residue is situated at the C-terminal position in the native protein. The polypeptide with 132 residues possessed N-acetylated residue at N-terminal position: therefore this polypeptide is located at the N-terminal position in the protein. The sum and account of each amino acid of the seven isolated peptides were compared to those of the intact protein: the sum of the seven peptides is 331 amino acid residues, whereas the whole protein contains 342 residues. The molecular weight of arginine kinase is revised and calculated on the basis of the present results (37, 687).
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis / Q000302:isolation & purification; D000818:Animals; D001120:Arginine; D002850:Chromatography, Gel; D002854:Chromatography, Paper; D003488:Cyanogen Bromide; D004590:Electrophoresis, Paper; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D008121:Nephropidae / Q000201:enzymology*; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D010770:Phosphotransferases / Q000032:analysis / Q000302:isolation & purification*
D000596:Amino Acids; D010455:Peptides; D001120:Arginine; D010770:Phosphotransferases; D003488:Cyanogen Bromide