Kinetics of hydrolysis of type I, II, and III collagens by the class I and II Clostridium histolyticum collagenases.
The kinetics of hydrolysis of rat tendon type I, bovine nasal septum type II, and human placental type III collagens by class I and class II Clostridium histolyticum collagenases (CHC) have been investigated. To facilitate this study, radioassays developed previously for the hydrolysis of these [3H]acetylated collagens by tissue collagenases have been adapted for use with the CHC. While the CHC are known to make multiple scissions in these collagens, the assays are shown to monitor the initial proteolytic events. The individual kinetic parameters kcat and KM have been determined for the hydrolysis of all three collagens by both class I and class II CHC. The specific activities of these CHC toward fibrillar type I and III collagens have also been measured. In contrast to human tissue collagenases, neither class of CHC exhibits a marked specificity toward any collagen type either in solution or in fibrillar form. The values of the kinetic parameters kcat and KM for the CHC are similar in magnitude to those of the human enzymes acting on their preferred substrates. Thus, the widely held view that the CHC are more potent collagenases is not strictly correct. As with the tissue collagenases, the local collagen structure at the cleavage sites is believed to play an important role in determining the rates of the reactions studied.
Mallya
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SK
Department of Chemistry, Florida State University, Tallahassee 32306.
Mookhtiar
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KA
Van Wart
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HE
D000107:Acetylation; D000818:Animals; D002417:Cattle; D003094:Collagen / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D046911:Macromolecular Substances; D003012:Microbial Collagenase / Q000378:metabolism*; D051381:Rats; D012680:Sensitivity and Specificity; D012967:Sodium Dodecyl Sulfate; D013696:Temperature; D014316:Tritium
D046911:Macromolecular Substances; D014316:Tritium; D012967:Sodium Dodecyl Sulfate; D003094:Collagen; D003012:Microbial Collagenase