Prevention of degradation of human polymorphonuclear leukocyte proteins by diisopropylfluorophosphate.

摘要: Proteases can complicate the characterization of proteins from cells, especially human polymorphonuclear leukocytes (PMN), which contain abundant neutral proteases. We tested ability agents to inhibit proteolysis, with special reference subunit polypeptides contractile actin, myosin, and actin-binding protein (ABP). Phenylmethylsulfonyl fluoride (PMSF), O-phenanthroline, EGTA, EDTA, N- ethylmaleimide, alone or in combinations, failed prevent extensive proteolysis PMN during solubilization cells dodecyl sulfate. These inhibitors also alpha-1-antitrypsin soybean trypsin inhibitor similarly could not homogenization cold isosomolar sucrose. Treatment greater than equal mM diisopropylfluorophosphate (DFP) prior markedly inhibited proteolysis. PMSF DFP were equally effective inhibiting extracts, suggesting that efficacy may result its permeation intact granules before barriers are disrupted by detergents homogenization. under conditions did affect their rate phagocytosis. recommend use future studies correlating functions structure PMN.