Purification of Tryptase From a Human Mast Cell Line

摘要: The neutral protease tryptase has been isolated from a human mast cell line, HMC-1. HMC-1 line was established the peripheral blood of patient with leukemia and maintained as continuously proliferating clones in vitro solid tumors nude mice. HMC-1-derived purified by sequential chromatography on Dowex 1, DEAE 5 PW, heparin-agarose. Purified an apparent molecular weight 150,000, determined sieve HPLC, but migrates doublet bands 32/35,000 SDS-PAGE gels. Maximal enzymatic activity observed at pH 8.5. Cleavage tosyl-L-arginine methyl ester inhibited dansyl-L-glutamyl-glycyl-L-arginine chloromethyl ketone 2 HCl, HgCl2, tosyl-L-lysine ketone, leupeptin, PMSF not benzamidine, aprotinin, tosyl-L-phenyl-alanine soybean trypsin inhibitor, plasma, ovomucoid or lima bean inhibitor. Microsequencing yielded amino terminal sequence that identical to previously reported for pituitary-derived tryptase.