Basic Study of Determination of Elastolytic Activity by Dialysis Method:I. Effect of Elastase on Soluble Elastin

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We previously reported a method of fluorometric determination of elastase activity after separating other protein and enzymatic reaction products by trichioroacetic acid.<BR>On the other hand, Substrate of soluble elastin (α-, β-elastin and elastase solubilized elastin) and other protein was not separated by the deproteinizing procedure. We improved above method separating substrate and enzymatic reaction products by dialysis method.<BR>As a result, we could get Km values of porcine pancreatic elastase, human pancreatic elastases 1 and 2 toward α-elastin, which were 0.51, 2, 84, 2.38 respectively and those toward β-elastin were 3.70, 2.24, 2.35 respectively. It may be suggested that porcine pancreatic elastase is tend to digest α-elastin rather β-elastin, but human pancreatic elastases 1 and 2 tend to digest similarly both elastins. Furthermore, we could find that elastase solubilized elastin was also digested by trypsin, chymotrypsin, collagenase and pronase. But insoluble elastin and α-, β-elastin as the substrate were not digested by the above enzymes.<BR>It has been suggested that insoluble elastin is primarily solubilized by elastase and then soluble elastin fragment is digested by other proteolytic enzymes.<BR>The elastase assay presented here should be usefull for the study of elastin metabolism under normal and various disease states.
Japan Society of Clinical Chemistryの論文