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

J. Biol. Chem., Vol. 280, Issue 10, 9160-9169, March 11, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/10/9160    most recent M409068200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Olivero, A. G. Articles by Kirchhofer, D. Search for Related Content PubMed PubMed Citation Articles by Olivero, A. G. Articles by Kirchhofer, D. Social Bookmarking                      What's this?

A Selective, Slow Binding Inhibitor of Factor VIIa Binds to a Nonstandard Active Site Conformation and Attenuates Thrombus Formation in Vivo^*

Alan G. Olivero, Charles Eigenbrot, Richard Goldsmith, Kirk Robarge, Dean R. Artis¶, John Flygare, Thomas Rawson, Daniel P. Sutherlin, Saloumeh Kadkhodayan||, Maureen Beresini, Linda O. Elliott, Geralyn G. DeGuzman**, David W. Banner, Mark Ultsch, Ulla Marzec¶¶, Stephen R. Hanson¶¶, Canio Refino**, Stuart Bunting**, and Daniel Kirchhofer**||||

From the Departments of Medicinal Chemistry, Protein Engineering, ^||Bioanalytical Research and Development, and ^**Physiology, Genentech, Inc., South San Francisco, California 94080, F. Hoffmann-La Roche, 4002 Basel, Switzerland, and Emory University, Atlanta, Georgia 30322

The serine protease factor VIIa (FVIIa) in complex with its cellular cofactor tissue factor (TF) initiates the blood coagulation reactions. TF·FVIIa is also implicated in thrombosis-related disorders and constitutes an appealing therapeutic target for treatment of cardiovascular diseases. To this end, we generated the FVIIa active site inhibitor G17905, which displayed great potency toward TF·FVIIa (K_i = 0.35 ± 0.11 nM). G17905 did not appreciably inhibit 12 of the 14 examined trypsin-like serine proteases, consistent with its TF·FVIIa-specific activity in clotting assays. The crystal structure of the FVIIa·G17905 complex provides insight into the molecular basis of the high selectivity. It shows that, compared with other serine proteases, FVIIa is uniquely equipped to accommodate conformational disturbances in the Gln²¹⁷–Gly²¹⁹ region caused by the ortho-hydroxy group of the inhibitor's aminobenzamidine moiety located in the S1 recognition pocket. Moreover, the structure revealed a novel, nonstandard conformation of FVIIa active site in the region of the oxyanion hole, a "flipped" Lys¹⁹²-Gly¹⁹³ peptide bond. Macromolecular substrate activation assays demonstrated that G17905 is a noncompetitive, slow-binding inhibitor. Nevertheless, G17905 effectively inhibited thrombus formation in a baboon arterio-venous shunt model, reducing platelet and fibrin deposition by 70% at 0.4 mg/kg + 0.1 mg/kg/min infusion. Therefore, the in vitro potency of G17905, characterized by slow binding kinetics, correlated with efficacious antithrombotic activity in vivo.

Received for publication, August 9, 2004 , and in revised form, December 23, 2004.

^* The Stanford Synchrotron Radiation Laboratory Structural Molecular Biology Program is supported by the Department of Energy, Office of Biological and Environmental Research and by the National Institutes of Health (NIH), National Center for Research Resources, Biomedical Technology Program, and NIGMS, NIH. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Present address: Plexxikon, Inc., Berkeley, CA 94710.

^¶¶ Present address: Dept. of Biomedical Engineering, OGI School of Science and Engineering, Oregon Health and Science University, Beaverton, OR 97006-8921.

^|||| To whom correspondence should be addressed: Dept. of Physiology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080. Tel.: 650-225-2134; Fax: 650-225-6327; E-mail: dak{at}gene.com.

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

This article has been cited by other articles:

				Y. Wu, C. Eigenbrot, W.-C. Liang, S. Stawicki, S. Shia, B. Fan, R. Ganesan, M. T. Lipari, and D. Kirchhofer Structural insight into distinct mechanisms of protease inhibition by antibodies PNAS, December 11, 2007; 104(50): 19784 - 19789. [Abstract] [Full Text] [PDF]

				R. A. Shirk and G. P. Vlasuk Inhibitors of Factor VIIa/Tissue Factor Arterioscler. Thromb. Vasc. Biol., September 1, 2007; 27(9): 1895 - 1900. [Abstract] [Full Text] [PDF]

				N. Mackman, R. E. Tilley, and N. S. Key Role of the Extrinsic Pathway of Blood Coagulation in Hemostasis and Thrombosis Arterioscler. Thromb. Vasc. Biol., August 1, 2007; 27(8): 1687 - 1693. [Abstract] [Full Text] [PDF]

				P. Moran, W. Li, B. Fan, R. Vij, C. Eigenbrot, and D. Kirchhofer Pro-urokinase-type Plasminogen Activator Is a Substrate for Hepsin J. Biol. Chem., October 13, 2006; 281(41): 30439 - 30446. [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]

				Y. Banerjee, J. Mizuguchi, S. Iwanaga, and R. M. Kini Hemextin AB Complex, a Unique Anticoagulant Protein Complex from Hemachatus haemachatus (African Ringhals Cobra) Venom That Inhibits Clot Initiation and Factor VIIa Activity J. Biol. Chem., December 30, 2005; 280(52): 42601 - 42611. [Abstract] [Full Text] [PDF]