Human lung tryptase. Purification and characterization.

摘要: Human lung tryptase, a mast cell-derived trypsin-like serine protease, has been isolated from whole human tissue obtained at autopsy. Increased yields this purification process have allowed extensive characterization of the enzyme. One critical steps in scheme is use linear heparin gradient to elute active material cellulose phosphate. Gel filtration studies 1.0 M NaCl yielded an apparent Mr = 135,000, and subsequent electrophoresis on sodium dodecyl sulfate-polyacrylamide gels demonstrated presence two species with 30,900 31,600. Enzymatic activity was sensitive concentrations above 0.05 only 50% 0.15 NaCl, decreasing 18% 0.6 NaCl. The effects synthetic natural inhibitors also studied, confirming enzyme's characteristics demonstrating that naturally occurring serum are incapable diminishing its activity. A complete amino acid analysis showed high tryptophan content. Lastly, antisera tryptase generated, immunological identity fractions investigated as well localization enzyme cell granule by immunohistochemical staining.