ISOLATION AND CHARACTERIZATION OF A 148 KDA PROTEIN POSSIBLY PARTICIPATING IN COLLAGEN-METABOLISM OF ABALONE ADDUCTOR MUSCLE

A 148 kDa protein which possibly participates in collagen metabolism w as isolated from the adductor muscle of the abalone Haliotis discus, b y a series of chromatographic procedures involving DEAE-Toyopearl 650 M ion exchange, TSK G3000SWG gel filtration, and Butyl-Toyopearl 650 M hydrophobic columns. Reactivity with collagen was measured as the abi lity to solubilize the film containing pepsin-solubilized collagen fro m the same animal. Active fractions of various purification steps solu bilized the film at 37 degrees C within 60 min. The purified protein c onsisted of a single polypeptide chain and the molecular weight was es timated by SDS-PAGE to be 148,000. This protein did not contain methio nine and half-cystine, and its N-terminal amino acid sequence differed from any sequences of known collagenases.