Purification of tryptase from a human mast cell line

J. H. Butterfield, D. A. Weiler, L. W. Hunt, S. R. Wynn, P. C. Roche

Research output: Contribution to journalArticle

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Abstract

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

Original languageEnglish (US)
Pages (from-to)409-419
Number of pages11
JournalJournal of Leukocyte Biology
Volume47
Issue number5
StatePublished - 1990

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Tryptases
Mast Cells
Cell Line
Trypsin Inhibitors
Mast-Cell Leukemia
Mercuric Chloride
Aprotinin
Ketones
Phenylalanine
Soybeans
Nude Mice
Lysine
Arginine
Chromatography
Polyacrylamide Gel Electrophoresis
Peptide Hydrolases
Clone Cells
Molecular Weight
Gels
High Pressure Liquid Chromatography

Keywords

  • leukemia
  • mast cell
  • mastocytomas
  • Neutral proteases

ASJC Scopus subject areas

  • Cell Biology

Cite this

Butterfield, J. H., Weiler, D. A., Hunt, L. W., Wynn, S. R., & Roche, P. C. (1990). Purification of tryptase from a human mast cell line. Journal of Leukocyte Biology, 47(5), 409-419.

Purification of tryptase from a human mast cell line. / Butterfield, J. H.; Weiler, D. A.; Hunt, L. W.; Wynn, S. R.; Roche, P. C.

In: Journal of Leukocyte Biology, Vol. 47, No. 5, 1990, p. 409-419.

Research output: Contribution to journalArticle

Butterfield, JH, Weiler, DA, Hunt, LW, Wynn, SR & Roche, PC 1990, 'Purification of tryptase from a human mast cell line', Journal of Leukocyte Biology, vol. 47, no. 5, pp. 409-419.
Butterfield JH, Weiler DA, Hunt LW, Wynn SR, Roche PC. Purification of tryptase from a human mast cell line. Journal of Leukocyte Biology. 1990;47(5):409-419.
Butterfield, J. H. ; Weiler, D. A. ; Hunt, L. W. ; Wynn, S. R. ; Roche, P. C. / Purification of tryptase from a human mast cell line. In: Journal of Leukocyte Biology. 1990 ; Vol. 47, No. 5. pp. 409-419.
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