Preliminary studies on cysteine and serine proteinase activities in inflamed human gingiva using different 7-amino-4-trifluoromethyl coumarin substrates and protease inhibitors.
The cysteine proteinases cathepsins B and L have collagenolytic potential and so have been implicated in connective-tissue breakdown in chronic periodontitis. Synthetic peptide substrates are often used to detect proteolytic enzymes. The action of homogenates of inflamed gingiva tissue against three such substrates of cathepsin B have been characterized here by protease inhibitors. Using the selective reagents ZPheAlaCHN2, BzValLysLysArgAFC, ZAlaArgArgAFC and ZPheArgAFC were susceptible to both cysteine and non-cysteine proteinase activity; the two types of enzymes had acidic and alkaline pH optima, respectively. The action of other inhibitors at acidic pH indicated the involvement of cathepsin B and, to a lesser extent, cathepsin L. The enzyme active at alkaline pH was a serine proteinase; it resembled glandular kallikrein in its inhibitor response and its ability to hydrolyse a fourth substrate, DValLeuArgAFC, but its greater reactivity with BzValLysLysArgAFC and ZAlaArgArgAFC was not consistent with kallikrein. ZPheArgAFC, though less sensitive than BzValLysLysArgAFC to cysteine proteinase action, was far less susceptible to hydrolysis by the serine proteinase and thus appears the best choice for selective assays of cathepsins B and L.
D000328:Adult; D003374:Coumarins / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors; D003978:Diazomethane / Q000031:analogs & derivatives / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005891:Gingivitis / Q000201:enzymology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D011480:Protease Inhibitors / Q000494:pharmacology*; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors; D013379:Substrate Specificity
D003374:Coumarins; D015853:Cysteine Proteinase Inhibitors; D011480:Protease Inhibitors; D015842:Serine Proteinase Inhibitors; C024918:benzyloxycarbonylphenylalanylalanine diazomethyl ketone; C055467:7-amino-4-trifluoromethylcoumarin; D003978:Diazomethane; D012697:Serine Endopeptidases; D003546:Cysteine Endopeptidases