30-kDa trypsin-like proteases in the plantar stratum corneum

The casein digestible proteases in human plantar stratum corneum were determined to be about 75-kDa, 30-kDa and 25-kDa in molecular weight by zymography. The enzymatic activity of the 75-kDa and 25-kDa proteases was specifically inhibited by chymostatin, which is an inhibitor of chymotrypsin-like serine proteases, and the proteases around 30-kDa were inhibited by leupeptin, a trypsin-like serine protease inhibitor. The enzymatic activity of all these proteases was inhibited by aprotinin. The 30-kDa trypsin-like proteases were heat-stable; their enzymatic activity still remained even after heating at 100 degrees C for 60 minutes. Their optimal pH was around 9, and the activity was higher in the outer part of the stratum corneum than in the inner part.