On the presence of proteolytic activity in glycosaminoglycan-degrading enzyme preparations.
A solid-phase protease assay has been used to screen different commercial preparations of glycosaminoglycan-degrading enzymes for the presence of proteolytic activity. Proteases cannot be detected in preparations of testicular hyaluronidase and of chondroitinase at the concentration used for histochemical purposes. Commercial Streptomyces hyaluronidase contains proteolytic contaminants detectable at the concentration used for histochemistry. At higher concentrations, all preparations appear to be contaminated with proteases. The results obtained using this assay suggest that addition of a mixture of proteinase inhibitors containing N-ethylmaleimide, EDTA, pepstatin, and phenylmethanesulfonylfluoride or soybean trypsin inhibitor has little effect on the proteolytic activity of the glycosaminoglycan-degrading enzyme preparations, irrespective of the pH used. Moreover, the use of EDTA in this mixture is questionable. This study also describes two testicular hyaluronidase preparations that may be particularly useful in functional studies of the living organism, as they are only slightly contaminated.
D000818:Animals; D002417:Cattle; D002808:Chondroitin Lyases / Q000378:metabolism; D004796:Clinical Enzyme Tests / Q000379:methods; D006025:Glycosaminoglycans / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D007202:Indicators and Reagents / Q000032:analysis; D008297:Male; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors; D012996:Solutions; D013737:Testis / Q000201:enzymology; D014357:Trypsin / Q000378:metabolism
D006025:Glycosaminoglycans; D007202:Indicators and Reagents; D011480:Protease Inhibitors; D012996:Solutions; D006821:Hyaluronoglucosaminidase; D010447:Peptide Hydrolases; D014357:Trypsin; D002808:Chondroitin Lyases