Urokinase-type plasminogen activator is a preferred substrate of the human epithelium serine protease tryptase /PRSS22
Authors:
- Nasa MorokawaNMNasa Morokawa
- G. William WongGWG. William Wong
- Andrea RossiARAndrea Rossi
- Mallur S. MadhusudhanMMMallur S. Madhusudhan
- Andrej ŠaliAŠAndrej Šali
- Yuko S. AskewYAYuko S. Askew
- Roberto AdachiRARoberto Adachi
- Gary A. SilvermanGSGary A. Silverman
- Steven A. KrilisSKSteven A. Krilis
- Richard L. StevensRSRichard L. Stevens
- 15 May 2005
- journal article
- Published by American Society of Hematology in Blood
- Vol. 105 (10) , 3893-3901
Abstract
Tryptase ϵ is a member of the chromosome 16p13.3 family of human serine proteases that is preferentially expressed by epithelial cells. Recombinant pro–tryptase ϵ was generated to understand how the exocytosed zymogen might be activated outside of the epithelial cell, as well as to address its possible role in normal and diseased states. Using expression/site-directed mutagenesis approaches, we now show that Lys20, Cys90, and Asp92 in the protease9s substrate-binding cleft regulate its enzymatic activity. We also show that Arg-1 in the propeptide domain controls its ability to autoactivate. In vitro studies revealed that recombinant tryptase ϵ possesses a restricted substrate specificity. Once activated, tryptase ϵ cannot be inhibited effectively by the diverse array of protease inhibitors present in normal human plasma. Moreover, this epithelium protease is not highly susceptible to α1-antitrypsin or secretory leukocyte protease inhibitor, which are present in the lung. Recombinant tryptase ϵ could not cleave fibronectin, vitronectin, laminin, single-chain tissue-type plasminogen activator, plasminogen, or any prominent serum protein. Nevertheless, tryptase ϵ readily converted single-chain pro–urokinase-type plasminogen activator (pro-uPA/scuPA) into its mature, enzymatically active protease. Tryptase ϵ also was able to induce pro-uPA–expressing smooth muscle cells to increase their migration through a basement membrane–like extracellular matrix. The ability to activate uPA in the presence of varied protease inhibitors suggests that tryptase ϵ plays a prominent role in fibrinolysis and other uPA-dependent reactions in the lung.Keywords
This publication has 49 references indexed in Scilit:
- Urokinase-deficient and urokinase receptor-deficient mice have impaired neutrophil antimicrobial activation in vitroJournal of Leukocyte Biology, 2004
- A novel serine protease predominately expressed in macrophagesBiochemical Journal, 2003
- The Crystal Structure of Human α1-Tryptase Reveals a Blocked Substrate-binding RegionJournal of Molecular Biology, 2002
- Rapid grid‐based construction of the molecular surface and the use of induced surface charge to calculate reaction field energies: Applications to the molecular systems and geometric objectsJournal of Computational Chemistry, 2001
- Structural Analysis of esp-1 Gene (PRSS 21)Biochemical and Biophysical Research Communications, 1999
- Cloning and Tissue Distribution of a Novel Serine Protease esp-1 from Human EosinophilsBiochemical and Biophysical Research Communications, 1998
- The Tryptase, Mouse Mast Cell Protease 7, Exhibits Anticoagulant Activity in Vivo and in Vitro Due to Its Ability to Degrade Fibrinogen in the Presence of the Diverse Array of Protease Inhibitors in PlasmaJournal of Biological Chemistry, 1997
- MOLMOL: A program for display and analysis of macromolecular structuresJournal of Molecular Graphics, 1996
- Comparative Protein Modelling by Satisfaction of Spatial RestraintsJournal of Molecular Biology, 1993
- Three-dimensional structure of the complex between pancreatic secretory trypsin inhibitor (Kazal type) and trypsinogen at 1.8 Å resolutionJournal of Molecular Biology, 1982