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Originally published In Press as doi:10.1074/jbc.M205868200 on August 22, 2002

J. Biol. Chem., Vol. 277, Issue 44, 41906-41915, November 1, 2002
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Biochemical and Functional Characterization of Human Transmembrane Tryptase (TMT)/Tryptase gamma
TMT IS AN EXOCYTOSED MAST CELL PROTEASE THAT INDUCES AIRWAY HYPERRESPONSIVENESS IN VIVO VIA AN INTERLEUKIN-13/INTERLEUKIN-4 RECEPTOR alpha /SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION (STAT) 6-DEPENDENT PATHWAY*,

Guang W. WongDagger §, Paul S. Foster, Shinsuke YasudaDagger , Jian C. Qi||, Surendran Mahalingam, Elizabeth A. MellorDagger , Gregory Katsoulotos||, Lixin LiDagger , Joshua A. BoyceDagger , Steven A. Krilis||, and Richard L. StevensDagger **

From the Dagger  Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 02115, the  Division of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, Canberra 0200, Australia, and the || Department of Medicine, University of New South Wales, and Department of Immunology, Allergy, and Infectious Disease, St. George Hospital, Kogarah, New South Wales 2217, Australia

Transmembrane tryptase (TMT)/tryptase gamma  is a membrane-bound serine protease stored in the secretory granules of human and mouse lung mast cells (MCs). We now show that TMT reaches the external face of the plasma membrane when MCs are induced to degranulate. Analysis of purified recombinant TMT revealed that it is a two-chain neutral protease. Thus, TMT is the only MC protease identified so far which retains its 18-residue propeptide when proteolytically activated. The genes that encode TMT and tryptase beta I reside on human chromosome 16p13.3. However, substrate specificity studies revealed that TMT and tryptase beta I are functionally distinct even though they are ~50% identical. Although TMT is rapidly inactivated by the human plasma serpin alpha 1-antitrypsin in vitro, administration of recombinant TMT (but not recombinant tryptase beta I) into the trachea of mice leads to airway hyperresponsiveness (AHR) and increased expression of interleukin (IL) 13. T cells also increase their expression of IL-13 mRNA when exposed to TMT in vitro. TMT is therefore a novel exocytosed surface mediator that can stimulate those cell types that are in close proximity. TMT induces AHR in normal mice but not in transgenic mice that lack signal transducer and activator of transcription (STAT) 6 or the alpha -chain of the cytokine receptor that recognizes both IL-4 and IL-13. Based on these data, we conclude that TMT is an exocytosed MC neutral protease that induces AHR in lungs primarily by activating an IL-13/IL-4Ralpha /STAT6-dependent pathway.

* This work was supported by grants from the National Health and Medical Research Council of Australia, the Charles Dana Foundation, and by Grants AI-23483, AI-31599, AI-48802, AI-52353, HL-36110, and HL-63284 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains a table of the transcript analysis of Jurkat T cells before and after exposure to TMT.

§ Pharmacia Allergy Research Fellow.

** To whom correspondence should be addressed: Dept. of Medicine, Brigham and Women's Hospital, Smith Bldg., Rm. 616B, 1 Jimmy Fund Way, Boston, MA 02115. Tel.: 617-525-1231; Fax: 617-525-1310; E-mail: rstevens@rics.bwh.harvard.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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