NDLI: Thrombin Interacts with Thrombomodulin, Protein C, and Thrombin-activatable Fibrinolysis Inhibitor via Specific and Distinct Domains

Content Provider	Journal of Biological Chemistry (JBC)
Author	Hall, Scott W. Nagashima, Mariko Zhao, Lei Morser, John Leung, Lawrence L. K.
Abstract	A collection of 56 purified thrombin mutants, in which 76 charged or polar surface residues on thrombin were mutated to alanine, was used to identify key residues mediating the interactions of thrombin with thrombomodulin (TM), protein C, and thrombin-activatable fibrinolysis inhibitor (TAFI). Comparison of protein C activation in the presence and absence of TM identified 11 residues mediating the thrombin-TM interaction (Lys21, Gln24, Arg62, Lys65, His66, Arg68, Thr69, Tyr71, Arg73, Lys77, Lys106). Three mutants (E25A, D51A, R89A/R93A/E94A) were found to have decreased ability to activate TAFI yet retained normal protein C activation, whereas three other mutants (R178A/R180A/D183A, E229A, R233A) had decreased ability to activate protein C but maintained normal TAFI activation. One mutant (W50A) displayed decreased activation of both substrates. Mapping of these functional residues on thrombin revealed that the 11 residues mediating the thrombin-TM interaction are all located in exosite I. Residues important in TAFI activation are located above the active-site cleft, whereas residues involved in protein C are located below the active-site cleft. In contrast to the extensive overlap of residues mediating TM binding and fibrinogen clotting, these data show that distinct domains in thrombin mediate its interactions with TM, protein C, and TAFI. These studies demonstrate that selective enzymatic properties of thrombin can be dissociated by site-directed mutagenesis.
Related Links	http://www.jbc.org/content/274/36/25510.abstract
Ending Page	25516
Starting Page	25510
Page Count	7
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	36
Volume Number	274
DOI	10.1074/jbc.274.36.25510
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	1999-09-03
Access Restriction	Open
Subject Keyword	Thrombomodulin (TM) Protein C (PC) Thrombin-activable fibrinolysis inhibitor (TAFI) Epidermal growth factor (EGF) 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonic acid (CHAPS) Phe-Pro-Arg-chloromethyl ketone (PPACK) Wild-type (WT) Activated protein C. CMV, cytomegalovirus (aPC) Chinese hamster ovary (CHO) PROTEIN CHEMISTRY AND STRUCTURE
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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