[Skin high-molecular-weight protease. 2. Biochemical properties].
The purification of High-Molecular-Weight Protease from Wistar Rat's skin was previously investigated and reported. Some properties of this enzyme were subsequently investigated. This enzyme showed high activities toward succynyl-leucyl-leucyl-valyl-tyrosine-methylcoumarinamide (SLLVT-MCA). The addition of 0.03% sodium dodecyl sulphate (SDS) stimulated enzyme activity to 2,000% of the initial activity. The pH optimum was pH 8.5 in the presence of 0.03% SDS, but pH 6.0-8.5 in the absence of SDS. This enzyme was inhibited by one of serine protease inhibitors, diisopropylfluorophosphate (DFP) strongly. It was also inhibited by some of SH protease inhibitors, N-ethylmaleimide (NEM), iodoacetamide (IA), and leupeptin. Interestingly, chymostatin, a serine and SH protease inhibitor also inhibited enzyme activity. These results show that both serine and cysteine++ residues are related to the enzyme activity of High-Molecular-Weight Protease.
Suga
Y
Y
Department of Dermatology, Juntendo University School of Medicine.
Yamada
H
H
Takamori
K
K
D000818:Animals; D003374:Coumarins / Q000378:metabolism; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012867:Skin / Q000201:enzymology*; D013379:Substrate Specificity
D003374:Coumarins; D009842:Oligopeptides; D011480:Protease Inhibitors; C059197:succinyl-leucyl-leucyl-valyl-tyrosyl-4-trifluoromethylcoumarin-7-amide; D010450:Endopeptidases