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 Higashi, S. Articles by Iwanaga, S. Search for Related Content PubMed PubMed Citation Articles by Higashi, S. Articles by Iwanaga, S. Social Bookmarking                      What's this?

Volume 271, Number 43, Issue of October 25, 1996 pp. 26569-26574
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Molecular Mechanism of Tissue Factor-mediated Acceleration of Factor VIIa Activity

(Received for publication, June 7, 1996, and in revised form, August 5, 1996)

Shouichi Higashi , Naomi Matsumoto and Sadaaki Iwanaga ^¶

From the  Department of Biology, Faculty of Science, and the ^¶ Department of Molecular Biology, Graduate School of Medical Science, Kyushu University 33, Fukuoka 812-81, Japan

The mechanism of the acceleration of the catalytic activity of factor VIIa (VIIa) in the presence of tissue factor (TF) was investigated. To explore the VIIa's site(s) that correlates with TF-mediated acceleration, zymogen VII, VIIa, and active site-modified VIIa were prepared, and dissociation constants (K_d) for their bindings to TF or soluble TF in solution were determined. We found that conversion of zymogen VII to VIIa led to an increase in affinity (G = 4.3-4.4 kJ/mol) for TFs. Dansyl-Glu-Gly-Arg chloromethyl ketone (DNS-EGRck) treatment of VIIa led to a further increase in the affinity (G = 7.3-12 kJ/mol). Neither removal of the Gla domain from VIIa nor truncation of the COOH-terminal membrane and cytoplasmic regions of TF affected the affinity enhanced after DNS-EGRck treatment of VIIa. Treatment of VIIa with (p-amidinophenyl)methanesulfonyl fluoride also enhanced its affinity for soluble TF, whereas treatment with 4-(2-aminoethyl)benzenesulfonyl fluoride, phenylmethylsulfonyl fluoride, or diisopropyl fluorophosphate had a slight effect on the affinity. On the other hand, DNS-EGRck and (p-amidinophenyl)methanesulfonyl fluoride treatments, but not diisopropyl fluorophosphate treatment, of VIIa led to protection of its -amino group of Ile-153 from carbamylation. Protection of the -amino group was consistent with formation of a critical salt bridge between Ile-153 and Asp-343 in the protease domain of VIIa. Therefore, TF may preferentially bind to the active conformational state of VIIa. When one assumes that free VIIa exists in equilibrium between minor active and dominant zymogen-like inactive conformational states, preferential binding of TF to the active state leads to a shift in equilibrium. We speculate that TF traps the active conformational state of VIIa and converts its zymogen-like state into an active state, thereby accelerating the VIIa activity.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. Toso, H. Zhu, and R. M. Camire The Conformational Switch from the Factor X Zymogen to Protease State Mediates Exosite Expression and Prothrombinase Assembly J. Biol. Chem., July 4, 2008; 283(27): 18627 - 18635. [Abstract] [Full Text] [PDF]

				K. D. Rand, M. D. Andersen, O. H. Olsen, T. J. D. Jorgensen, H. Ostergaard, O. N. Jensen, H. R. Stennicke, and E. Persson The Origins of Enhanced Activity in Factor VIIa Analogs and the Interplay between Key Allosteric Sites Revealed by Hydrogen Exchange Mass Spectrometry J. Biol. Chem., May 9, 2008; 283(19): 13378 - 13387. [Abstract] [Full Text] [PDF]

				S. P. Bajaj, A. E. Schmidt, S. Agah, M. S. Bajaj, and K. Padmanabhan High Resolution Structures of p-Aminobenzamidine- and Benzamidine-VIIa/Soluble Tissue Factor: UNPREDICTED CONFORMATION OF THE 192-193 PEPTIDE BOND AND MAPPING OF Ca2+, Mg2+, Na+, AND Zn2+ SITES IN FACTOR VIIa J. Biol. Chem., August 25, 2006; 281(34): 24873 - 24888. [Abstract] [Full Text] [PDF]

				K. Yamamoto, S. Higashi, M. Kioi, J. Tsunezumi, K. Honke, and K. Miyazaki Binding of Active Matrilysin to Cell Surface Cholesterol Sulfate Is Essential for Its Membrane-associated Proteolytic Action and Induction of Homotypic Cell Adhesion J. Biol. Chem., April 7, 2006; 281(14): 9170 - 9180. [Abstract] [Full Text] [PDF]

				H. R. Maun, C. Eigenbrot, H. Raab, D. Arnott, L. Phu, S. Bullens, and R. A. Lazarus Disulfide locked variants of factor VIIa with a restricted {beta}-strand conformation have enhanced enzymatic activity Protein Sci., May 1, 2005; 14(5): 1171 - 1180. [Abstract] [Full Text] [PDF]

				K. Soejima, M. Yuguchi, J. Mizuguchi, K. Tomokiyo, T. Nakashima, T. Nakagaki, and S. Iwanaga The 99 and 170 Loop-modified Factor VIIa Mutants Show Enhanced Catalytic Activity without Tissue Factor J. Biol. Chem., December 6, 2002; 277(50): 49027 - 49035. [Abstract] [Full Text] [PDF]

				R. M. Camire Prothrombinase Assembly and S1 Site Occupation Restore the Catalytic Activity of FXa Impaired by Mutation at the Sodium-binding Site J. Biol. Chem., September 27, 2002; 277(40): 37863 - 37870. [Abstract] [Full Text] [PDF]

				A. Iakhiaev, U. R. Pendurthi, and L. V. M. Rao Active Site Blockade of Factor VIIa Alters Its Intracellular Distribution J. Biol. Chem., November 30, 2001; 276(49): 45895 - 45901. [Abstract] [Full Text] [PDF]

				E. Persson, M. Kjalke, and O. H. Olsen Rational design of coagulation factor VIIa variants with substantially increased intrinsic activity PNAS, October 31, 2001; (2001) 241339498. [Abstract] [Full Text] [PDF]

				A. C. W. Pike, A. M. Brzozowski, S. M. Roberts, O. H. Olsen, and E. Persson Structure of human factor VIIa and its implications for the triggering of blood coagulation PNAS, August 3, 1999; 96(16): 8925 - 8930. [Abstract] [Full Text] [PDF]

				F. Werner, T. M. Razzaq, and V. Ellis Tissue Plasminogen Activator Binds to Human Vascular Smooth Muscle Cells by a Novel Mechanism. EVIDENCE FOR A RECIPROCAL LINKAGE BETWEEN INHIBITION OF CATALYTIC ACTIVITY AND CELLULAR BINDING J. Biol. Chem., July 30, 1999; 274(31): 21555 - 21561. [Abstract] [Full Text] [PDF]

				C. D. McCallum, B. Su, P. F. Neuenschwander, J. H. Morrissey, and A. E. Johnson Tissue Factor Positions and Maintains the Factor VIIa Active Site Far above the Membrane Surface Even in the Absence of the Factor VIIa Gla Domain. A FLUORESCENCE RESONANCE ENERGY TRANSFER STUDY J. Biol. Chem., November 28, 1997; 272(48): 30160 - 30166. [Abstract] [Full Text] [PDF]

				S. Higashi, N. Matsumoto, and S. Iwanaga Conformation of Factor VIIa Stabilized by a Labile Disulfide Bond (Cys-310-Cys-329) in the Protease Domain Is Essential for Interaction with Tissue Factor J. Biol. Chem., October 10, 1997; 272(41): 25724 - 25730. [Abstract] [Full Text] [PDF]

				C. D. Dickinson and W. Ruf Active Site Modification of Factor VIIa Affects Interactions of the Protease Domain with Tissue Factor J. Biol. Chem., August 8, 1997; 272(32): 19875 - 19879. [Abstract] [Full Text] [PDF]

				B. B. Sorensen, E. Persson, P.-O. Freskgard, M. Kjalke, M. Ezban, T. Williams, and L. V. M. Rao Incorporation of an Active Site Inhibitor in Factor VIIa Alters the Affinity for Tissue Factor J. Biol. Chem., May 2, 1997; 272(18): 11863 - 11868. [Abstract] [Full Text] [PDF]

				R. J. Petrovan and W. Ruf Residue Met156 Contributes to the Labile Enzyme Conformation of Coagulation Factor VIIa J. Biol. Chem., February 23, 2001; 276(9): 6616 - 6620. [Abstract] [Full Text] [PDF]

				K. Soejima, J. Mizuguchi, M. Yuguchi, T. Nakagaki, S. Higashi, and S. Iwanaga Factor VIIa Modified in the 170 Loop Shows Enhanced Catalytic Activity but Does Not Change the Zymogen-like Property J. Biol. Chem., May 11, 2001; 276(20): 17229 - 17235. [Abstract] [Full Text] [PDF]

				E. Persson, M. Kjalke, and O. H. Olsen Rational design of coagulation factor VIIa variants with substantially increased intrinsic activity PNAS, November 20, 2001; 98(24): 13583 - 13588. [Abstract] [Full Text] [PDF]