[The purification of high molecular weight protease from the rat's skin]

In this study, the purification of a high molecular weight protease from Wistar Rats' skin was investigated. A high molecular weight protease from Wistar Rats' skin was purified by successive (NH4)2SO4 fractionation, Phenylsepharose CL-4B, and HPLC gel filtration chromatographies. The activity of the enzyme was determined using a synthetic substrate, succinyl-leucyl-leucyl-valyl-tyrosine methylcoumarinamide (SLLVT-MCA). The molecular weight of the purified enzyme was estimated to be 750,000 by gel filtration. With SDS-PAGE the purified enzyme was dissociated into several bands of which the majority fell into the range of 20,100-36,000 daltons. These findings suggest that high molecular weight protease in rat's skin consists of various subunits.