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Human Lung Tryptase Purification And Characterization, Timothy J. Smith

Electronic Theses and Dissertations

Human lung tryptase (HLT), a mast cell derived trypsin-like enzyme, was isolated from whole human lung tissue obtained at autopsy. Increased yields from this purification process allowed extensive characterization of the enzyme. One of the critical steps in the purification scheme was the use of a linear heparin gradient to elute active material from cellulose phosphate. Gel filtration studies in 1.0 M NaCl yielded an apparent M(,r) of 135,000, and subsequent electrophoresis on sodium dodecyl sulfate-polyacrylamide gels demonstrated the presence of two active species with apparent M(,r) = 30,900 and 31,600. Enzymatic activity was sensitive to NaCl concentrations above 0.05 …