| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 263, Issue 12, 5714-5723, 04, 1988
S Krishnaswamy and KG Mann
The analysis of free sulfhydryl groups in factor Va using dithiobis-
(nitrobenzoic acid) (DTNB) indicated the presence of one accessible thiol
in each of the two subunits of the cofactor. Intact factor Va contained one
readily accessible sulfhydryl group under native conditions and
approximately two such groups after denaturation. A comparison of the rate
of modification of the accessible thiol in factor Va under native
conditions to those observed with the isolated subunits indicated that the
thiol present in component D of the cofactor was readily accessible to
reaction with DTNB. Factor Va was reacted with the sulfhydryl-directed
fluorophore N-(1-pyrene)maleimide, resulting in the concomitant loss of the
accessible thiol with no detectable alteration in the activity of the
cofactor. This fluorescent derivative of factor Va (Pyr-Va) was used to
examine the binding of factor Va to phospholipid vesicles by fluorescence
polarization. Fluorescence polarization of the pyrene moiety increased
saturably when Pyr-Va was titrated with increasing concentrations of
vesicles composed of phosphatidylcholine and phosphatidylserine (PS).
Systematic analysis of the binding of Pyr-Va to PCPS (75%
phosphatidylcholine, 25% PS) indicated that the binding interaction was
characterized by a dissociation constant of 2.7 x 10(-9) M with 42 mol of
PCPS bound per mol of Va at saturation. The data obtained by varying the PS
content of the vesicles are consistent with the interpretation that the Va-
combining site on the vesicle surface is composed of a discrete number of
PS molecules. The binding of Pyr-Va to PCPS was independent of added
calcium ion and could be reversed by the addition of unlabeled Va or
isolated component E but not by component D. Analysis of the displacement
curves indicated that native factor Va or isolated component E and Pyr-Va
mutually excluded each other on the vesicle surface with identical
affinities. Competition experiments conducted using component E digested by
factor Xa or the isolated derivative peptides indicated that the cleavage
of component E by factor Xa had no effect on the PCPS binding properties of
this subunit. Further, the data obtained with the isolated peptides suggest
that the lipid-binding domain of component E is present in the
amino-terminal region of this subunit.
The binding of factor Va to phospholipid vesicles
Department of Biochemistry, University of Vermont, Burlington 05405.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  S. B. Jeimy, R. A. Woram, N. Fuller, M. A. Quinn-Allen, G. A. F. Nicolaes, B. Dahlback, W. H. Kane, and C. P. M. Hayward Identification of the MMRN1 Binding Region within the C2 Domain of Human Factor V J. Biol. Chem., December 3, 2004; 279(49): 51466 - 51471. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yegneswaran, R. M. Mesters, J. A. Fernandez, and J. H. Griffin Prothrombin Residues 473-487 Contribute to Factor Va Binding in the Prothrombinase Complex J. Biol. Chem., November 19, 2004; 279(47): 49019 - 49025. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. E. Adams, M. F. Hockin, K. G. Mann, and S. J. Everse The crystal structure of activated protein C-inactivated bovine factor Va: Implications for cofactor function PNAS, June 15, 2004; 101(24): 8918 - 8923. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. S. Boskovic, T. Troxler, and S. Krishnaswamy Active Site-independent Recognition of Substrates and Product by Bovine Prothrombinase: A FLUORESCENCE RESONANCE ENERGY TRANSFER STUDY J. Biol. Chem., May 14, 2004; 279(20): 20786 - 20793. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Toso and R. M. Camire Removal of B-domain Sequences from Factor V Rather than Specific Proteolysis Underlies the Mechanism by Which Cofactor Function Is Realized J. Biol. Chem., May 14, 2004; 279(20): 21643 - 21650. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Orfeo, N. Brufatto, M. E. Nesheim, H. Xu, S. Butenas, and K. G. Mann The Factor V Activation Paradox J. Biol. Chem., May 7, 2004; 279(19): 19580 - 19591. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kalafatis, D. O. Beck, and K. G. Mann Structural Requirements for Expression of Factor Va Activity J. Biol. Chem., August 29, 2003; 278(35): 33550 - 33561. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. G. Mann and M. Kalafatis Factor V: a combination of Dr Jekyll and Mr Hyde Blood, January 1, 2003; 101(1): 20 - 30. [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]
|
|
|
|
|
|
R. Majumder, G. Weinreb, X. Zhai, and B. R. Lentz Soluble Phosphatidylserine Triggers Assembly in Solution of a Prothrombin-activating Complex in the Absence of a Membrane Surface J. Biol. Chem., August 9, 2002; 277(33): 29765 - 29773. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. K. Buddai, L. Toulokhonova, P. W. Bergum, G. P. Vlasuk, and S. Krishnaswamy Nematode Anticoagulant Protein c2 Reveals a Site on Factor Xa That Is Important for Macromolecular Substrate Binding to Human Prothrombinase J. Biol. Chem., July 12, 2002; 277(29): 26689 - 26698. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Wilkens and S. Krishnaswamy The Contribution of Factor Xa to Exosite-dependent Substrate Recognition by Prothrombinase J. Biol. Chem., March 8, 2002; 277(11): 9366 - 9374. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. E. Gilbert, R. J. Kaufman, A. A. Arena, H. Miao, and S. W. Pipe Four Hydrophobic Amino Acids of the Factor VIII C2 Domain Are Constituents of Both the Membrane-binding and von Willebrand Factor-binding Motifs J. Biol. Chem., February 15, 2002; 277(8): 6374 - 6381. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-S. Mao, C. T. Przysiecki, J. A. Krueger, C. M. Cooper, S. D. Lewis, J. Joyce, C. Lellis, V. M. Garsky, M. Sardana, and J. A. Shafer Selective Inhibition of Factor Xa in the Prothrombinase Complex by the Carboxyl-terminal Domain of Antistasin J. Biol. Chem., November 13, 1998; 273(46): 30086 - 30091. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Betz and S. Krishnaswamy Regions Remote from the Site of Cleavage Determine Macromolecular Substrate Recognition by the Prothrombinase Complex J. Biol. Chem., April 24, 1998; 273(17): 10709 - 10718. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kalafatis Identification and Partial Characterization of Factor Va Heavy Chain Kinase from Human Platelets J. Biol. Chem., April 3, 1998; 273(14): 8459 - 8466. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. D. Spiess Endothelial Cell-Blood Interface Actions and the Procoagulant Response Seminars in Cardiothoracic and Vascular Anesthesia, November 1, 1997; 1(4): 288 - 294. [Abstract] [PDF]
|
|
|
|
 |
|
 M. F. Hockin, M. Kalafatis, M. Shatos, and K. G. Mann Protein C Activation and Factor Va Inactivation on Human Umbilical Vein Endothelial Cells Arterioscler. Thromb. Vasc. Biol., November 1, 1997; 17(11): 2765 - 2775. [Abstract] [Full Text]
|
|
|
|
|
|
K. G. Mann, M. F. Hockin, K. J. Begin, and M. Kalafatis Activated Protein C Cleavage of Factor Va Leads to Dissociation of the A2 Domain J. Biol. Chem., August 15, 1997; 272(33): 20678 - 20683. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kalafatis and K. G. Mann Factor VLeiden and Thrombophilia Arterioscler. Thromb. Vasc. Biol., April 1, 1997; 17(4): 620 - 627. [Full Text]
|
|
|
|
|
|
E. L.G. Pryzdial and G. E. Kessler Kinetics of Blood Coagulation Factor Xaalpha Autoproteolytic Conversion to Factor Xabeta . EFFECT ON INHIBITION BY ANTITHROMBIN, PROTHROMBINASE ASSEMBLY, AND ENZYME ACTIVITY J. Biol. Chem., July 12, 1996; 271(28): 16621 - 16626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. P. M. Hayward, E. Furmaniak-Kazmierczak, A.-M. Cieutat, J. C. Moore, D. F. Bainton, M. E. Nesheim, J. G. Kelton, and G. Côté Factor V Is Complexed with Multimerin in Resting Platelet Lysates and Colocalizes with Multimerin in Platelet alpha-Granules J. Biol. Chem., August 18, 1995; 270(33): 19217 - 19224. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. R. Zeibdawi and E. L. G. Pryzdial Mechanism of Factor Va Inactivation by Plasmin. LOSS OF A2 AND A3 DOMAINS FROM A Ca2+-DEPENDENT COMPLEX OF FRAGMENTS BOUND TO PHOSPHOLIPID J. Biol. Chem., June 1, 2001; 276(23): 19929 - 19936. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. S. Boskovic and S. Krishnaswamy Exosite Binding Tethers the Macromolecular Substrate to the Prothrombinase Complex and Directs Cleavage at Two Spatially Distinct Sites J. Biol. Chem., December 1, 2000; 275(49): 38561 - 38570. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
