Purification and characterization of a soluble endopeptidase from rat bone.
A soluble endopeptidase was purified from rat bone using ammonium sulphate precipitation followed by Fast Protein Liquid Chromatography on gel, anion exchange, and chromatofocusing columns. The enzyme was silver stain-pure (SDS-PAGE) and its apparent molecular weight was 60,000. In general, the enzyme favored the hydrolysis of the X-Y bond in substrates with the sequence of -Pro-X-Y-Pro-, where either X or Y (or both) were hydrophobic residues. The presence of imino acid residues near the scissile bond favored hydrolysis. The enzyme was strongly inactivated by metal chelating agents and p-hydroxymercuribenzoic acid, indicating that SH-groups may be necessary for full activity of the enzyme and that the enzyme may be a metallopeptidase.
Bollen
A M
AM
Department of Biologic and Material Sciences, School of Dentistry, University of Michigan, Ann Arbor 48109.
Makinen
K K
KK
D000595:Amino Acid Sequence; D000818:Animals; D001842:Bone and Bones / Q000201:enzymology*; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility; D013379:Substrate Specificity
D009842:Oligopeptides; D010450:Endopeptidases