Detection and characterization of epidermal proteinases by polyacrylamide gel electrophoresis.
Electrophoresis of cornified cell extracts of 2-day-old rats, using SDS polyacrylamide gels copolymerized with alpha-casein or gelatin with or without plasminogen, was performed. Both Tris-HCl buffer soluble protein and KSCN solubilized protein contained a number of hydrolases for alpha-casein and/or gelatin, but PA (mol. wts 57 and 50K) was found only in the KSCN extract. The pH dependency, substrate specificity and mol. wt of plasminogen-independent proteinases were comparatively determined and DFP inhibition tested. This simple technique helped to identify the presence of several proteinases and to characterize them in partially purified epidermal extracts.
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010960:Plasminogen Activators / Q000302:isolation & purification; D011480:Protease Inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D012867:Skin / Q000201:enzymology*; D013379:Substrate Specificity
D011480:Protease Inhibitors; D010450:Endopeptidases; D010960:Plasminogen Activators