HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Edwards, K. M. Articles by Trapani, J. A. Search for Related Content PubMed PubMed Citation Articles by Edwards, K. M. Articles by Trapani, J. A. Social Bookmarking                      What's this?

J Biol Chem, Vol. 274, Issue 43, 30468-30473, October 22, 1999

The Human Cytotoxic T Cell Granule Serine Protease Granzyme H Has Chymotrypsin-like (Chymase) Activity and Is Taken Up into Cytoplasmic Vesicles Reminiscent of Granzyme B-containing Endosomes

Kirsten M. Edwards, Chih-Min Kam^§, James C. Powers^§, and Joseph A. Trapani

From the  The John Connell Laboratory, The Austin Research Institute, Studley Road, Heidelberg, 3084, Australia and the ^§ School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia 30332-0400

Serine proteases (granzymes) contained within the cytoplasmic granules of cytotoxic T cells and natural killer cells play a variety of roles including the induction of target cell apoptosis, breakdown of extracellular matrix proteins and induction of cytokine secretion by bystander leukocytes. Different granzymes display proteolytic specificities that mimic the activities of trypsin or chymotrypsin, or may cleave substrates at acidic ("Asp-ase") or at long unbranched amino acids such as Met ("Met-ase"). Here, we report that recombinant granzyme H has chymotrypsin-like (chymase) activity, the first report of a human granzyme with this proteolytic specificity. Recombinant 32-kDa granzyme H expressed in the baculovirus vector pBacPAK8 was secreted from Sf21 cells and recovered by Ni-affinity chromatography, using a poly-His tag encoded at the predicted carboxyl terminus of full-length granzyme H cDNA. The granzyme H efficiently cleaved Suc-Phe-Leu-Phe-SBzl (v = 185 nM/s at [S] = 0.217 mM) and also hydrolyzed Boc-Ala-Ala-X-SBzl (X = Phe, Tyr, Met, Nle, or Nva) with slower rates but had little tryptase or Asp-ase activity. Enzymatic activity was inhibited completely by 0.1 mM 3,4-dichloroisocoumarin and 84% by 1.0 mM phenylmethylsulfonyl fluoride. Fluoresceinated granzyme H was internalized in a temperature-dependent manner by Jurkat cells into endosome-like vesicles, suggesting that it can bind to cell surface receptors similar to those that bind granzyme B. This suggests a hitherto unsuspected intracellular function for granzyme H.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				G. H. Caughey, J. Beauchamp, D. Schlatter, W. W. Raymond, N. N. Trivedi, D. Banner, H. Mauser, and J. Fingerle Guinea Pig Chymase Is Leucine-specific: A NOVEL EXAMPLE OF FUNCTIONAL PLASTICITY IN THE CHYMASE/GRANZYME FAMILY OF SERINE PEPTIDASES J. Biol. Chem., May 16, 2008; 283(20): 13943 - 13951. [Abstract] [Full Text] [PDF]

				R Tsuru, H Kondo, Y Hojo, M Gama, O Mizuno, T Katsuki, K Shimada, M Kikuchi, and T Yashiro Increased granzyme B production from peripheral blood mononuclear cells in patients with acute coronary syndrome Heart, March 1, 2008; 94(3): 305 - 310. [Abstract] [Full Text] [PDF]

				E. Fellows, S. Gil-Parrado, D. E. Jenne, and F. C. Kurschus Natural killer cell-derived human granzyme H induces an alternative, caspase-independent cell-death program Blood, July 15, 2007; 110(2): 544 - 552. [Abstract] [Full Text] [PDF]

				V. R. Sutton, N. J. Waterhouse, K. A. Browne, K. Sedelies, A. Ciccone, D. Anthony, A. Koskinen, A. Mullbacher, and J. A. Trapani Residual active granzyme B in cathepsin C-null lymphocytes is sufficient for perforin-dependent target cell apoptosis J. Cell Biol., February 12, 2007; 176(4): 425 - 433. [Abstract] [Full Text] [PDF]

				C. Adrain, P. J. Duriez, G. Brumatti, P. Delivani, and S. J. Martin The Cytotoxic Lymphocyte Protease, Granzyme B, Targets the Cytoskeleton and Perturbs Microtubule Polymerization Dynamics J. Biol. Chem., March 24, 2006; 281(12): 8118 - 8125. [Abstract] [Full Text] [PDF]

				L. I. Pao, N. Sumaria, J. M. Kelly, S. v. Dommelen, E. Cretney, M. E. Wallace, D. A. Anthony, A. P. Uldrich, D. I. Godfrey, J. M. Papadimitriou, et al. Functional Analysis of Granzyme M and Its Role in Immunity to Infection J. Immunol., September 1, 2005; 175(5): 3235 - 3243. [Abstract] [Full Text] [PDF]

				S. Mahrus, W. Kisiel, and C. S. Craik Granzyme M Is a Regulatory Protease That Inactivates Proteinase Inhibitor 9, an Endogenous Inhibitor of Granzyme B J. Biol. Chem., December 24, 2004; 279(52): 54275 - 54282. [Abstract] [Full Text] [PDF]

				K. A. Lindstedt and P. T. Kovanen Proteolysis of Pericellular Matrix: A Process Linking Inflammation to Plaque Destabilization and Rupture Arterioscler. Thromb. Vasc. Biol., December 1, 2004; 24(12): 2205 - 2206. [Full Text] [PDF]

				K. A. Lindstedt, M. J. Leskinen, and P. T. Kovanen Proteolysis of the Pericellular Matrix: A Novel Element Determining Cell Survival and Death in the Pathogenesis of Plaque Erosion and Rupture Arterioscler. Thromb. Vasc. Biol., August 1, 2004; 24(8): 1350 - 1358. [Abstract] [Full Text] [PDF]

				J. A. MacLean II, R. M. Roberts, and J. A. Green Atypical Kunitz-Type Serine Proteinase Inhibitors Produced by the Ruminant Placenta Biol Reprod, August 1, 2004; 71(2): 455 - 463. [Abstract] [Full Text] [PDF]

				K. A. Sedelies, T. J. Sayers, K. M. Edwards, W. Chen, D. G. Pellicci, D. I. Godfrey, and J. A. Trapani Discordant Regulation of Granzyme H and Granzyme B Expression in Human Lymphocytes J. Biol. Chem., June 18, 2004; 279(25): 26581 - 26587. [Abstract] [Full Text] [PDF]

				J. M. Kelly, N. J. Waterhouse, E. Cretney, K. A. Browne, S. Ellis, J. A. Trapani, and M. J. Smyth Granzyme M Mediates a Novel Form of Perforin-dependent Cell Death J. Biol. Chem., May 21, 2004; 279(21): 22236 - 22242. [Abstract] [Full Text] [PDF]

				O Ozdemir and S Savasan The role of mast cells in bone marrow diseases J. Clin. Pathol., January 1, 2004; 57(1): 108 - 109. [Full Text]

				H. Johnson, L. Scorrano, S. J. Korsmeyer, and T. J. Ley Cell death induced by granzyme C Blood, April 15, 2003; 101(8): 3093 - 3101. [Abstract] [Full Text] [PDF]

				C. E. Hirst, M. S. Buzza, C. H. Bird, H. S. Warren, P. U. Cameron, M. Zhang, P. G. Ashton-Rickardt, and P. I. Bird The Intracellular Granzyme B Inhibitor, Proteinase Inhibitor 9, Is Up-Regulated During Accessory Cell Maturation and Effector Cell Degranulation, and Its Overexpression Enhances CTL Potency J. Immunol., January 15, 2003; 170(2): 805 - 815. [Abstract] [Full Text] [PDF]

				S. Blesson, J. Thiery, C. Gaudin, R. Stancou, J.-P. Kolb, J.-L. Moreau, J. Theze, F. Mami-Chouaib, and S. Chouaib Analysis of the mechanisms of human cytotoxic T lymphocyte response inhibition by NO Int. Immunol., October 1, 2002; 14(10): 1169 - 1178. [Abstract] [Full Text] [PDF]

				C. E. Hirst, M. S. Buzza, V. R. Sutton, J. A. Trapani, K. L. Loveland, and P. I. Bird Perforin-independent expression of granzyme B and proteinase inhibitor 9 in human testis and placenta suggests a role for granzyme B-mediated proteolysis in reproduction Mol. Hum. Reprod., December 1, 2001; 7(12): 1133 - 1142. [Abstract] [Full Text] [PDF]

				M. J. Smyth, J. M. Kelly, V. R. Sutton, J. E. Davis, K. A. Browne, T. J. Sayers, and J. A. Trapani Unlocking the secrets of cytotoxic granule proteins J. Leukoc. Biol., July 1, 2001; 70(1): 18 - 29. [Abstract] [Full Text] [PDF]

				T. J. Sayers, A. D. Brooks, J. M. Ward, T. Hoshino, W. E. Bere, G. W. Wiegand, J. M. Kelley, and M. J. Smyth The Restricted Expression of Granzyme M in Human Lymphocytes J. Immunol., January 15, 2001; 166(2): 765 - 771. [Abstract] [Full Text] [PDF]