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

J. Biol. Chem., Vol. 281, Issue 51, 39194-39204, December 22, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/51/39194    most recent M604482200v2 M604482200v1 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 Bukys, M. A. Articles by Kalafatis, M. Search for Related Content PubMed PubMed Citation Articles by Bukys, M. A. Articles by Kalafatis, M. Social Bookmarking                      What's this?

A Control Switch for Prothrombinase

CHARACTERIZATION OF A HIRUDIN-LIKE PENTAPEPTIDE FROM THE COOH TERMINUS OF FACTOR Va HEAVY CHAIN THAT REGULATES THE RATE AND PATHWAY FOR PROTHROMBIN ACTIVATION^*

Michael A. Bukys, Paul Y. Kim, Michael E. Nesheim^||, and Michael Kalafatis^¶1

From the Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115, the Departments of Biochemistry and ^||Medicine, Queen's University, Kingston, Ontario K7L 3N6, Canada, and the ^¶Department of Molecular Cardiology, The Lerner Research Institute, The Cleveland Clinic, Cleveland, Ohio 44195

Membrane-bound factor Xa alone catalyzes prothrombin activation following initial cleavage at Arg²⁷¹ and prethrombin 2 formation (pre2 pathway). Factor Va directs prothrombin activation by factor Xa through the meizothrombin pathway, characterized by initial cleavage at Arg³²⁰ (meizo pathway). We have shown previously that a pentapeptide encompassing amino acid sequence 695–699 from the COOH terminus of the heavy chain of factor Va (Asp-Tyr-Asp-Tyr-Gln, DYDYQ) inhibits prothrombin activation by prothrombinase in a competitive manner with respect to substrate. To understand the mechanism of inhibition of thrombin formation by DYDYQ, we have studied prothrombin activation by gel electrophoresis. Titration of plasma-derived prothrombin activation by prothrombinase, with increasing concentrations of peptide, resulted in complete inhibition of the meizo pathway. However, thrombin formation still occurred through the pre2 pathway. These data demonstrate that the peptide preferentially inhibits initial cleavage of prothrombin by prothrombinase at Arg³²⁰. These findings were corroborated by studying the activation of recombinant mutant prothrombin molecules rMZ-II (R155A/R284A/R271A) and rP2-II (R155A/R284A/R320A) which can be only cleaved at Arg³²⁰ and Arg²⁷¹, respectively. Cleavage of rMZ-II by prothrombinase was completely inhibited by low concentrations of DYDYQ, whereas high concentrations of pentapeptide were required to inhibit cleavage of rP2-II. The pentapeptide also interfered with prothrombin cleavage by membrane-bound factor Xa alone in the absence of factor Va increasing the rate for cleavage at Arg²⁷¹ of plasma-derived prothrombin or rP2-II. Our data demonstrate that pentapeptide DYDYQ has opposing effects on membrane-bound factor Xa for prothrombin cleavage, depending on the incorporation of factor Va in prothrombinase.

Received for publication, May 10, 2006 , and in revised form, October 3, 2006.

^* This work was supported by a predoctoral fellowship from the Cellular and Molecular Medicine Specialization at Cleveland State University (to M. A. B.), United States Public Health Services Grant HL-46703-6 from the National Institutes of Health (to M. E. N.), and NHLBI Grant R01 HL-73343 from the National Institutes of Health (to M. K.). 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.

¹ To whom correspondence should be addressed: Dept. of Chemistry, Cleveland State University, 2351 Euclid Ave., Science and Research Center SR370, Cleveland, OH 44115. Tel.: 216-687-2460; Fax: 216-687-9298; E-mail: m.kalafatis{at}csuohio.edu.

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

This article has been cited by other articles:

				S. Tran, E. Norstrom, and B. Dahlback Effects of Prothrombin on the Individual Activated Protein C-mediated Cleavages of Coagulation Factor Va J. Biol. Chem., March 14, 2008; 283(11): 6648 - 6655. [Abstract] [Full Text] [PDF]

				P. Y. Kim and M. E. Nesheim Further Evidence for Two Functional Forms of Prothrombinase Each Specific for Either of the Two Prothrombin Activation Cleavages J. Biol. Chem., November 9, 2007; 282(45): 32568 - 32581. [Abstract] [Full Text] [PDF]

				A. J. Gale, S. Yegneswaran, X. Xu, J.-L. Pellequer, and J. H. Griffin Characterization of a Factor Xa Binding Site on Factor Va near the Arg-506 Activated Protein C Cleavage Site J. Biol. Chem., July 27, 2007; 282(30): 21848 - 21855. [Abstract] [Full Text] [PDF]