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Human Alpha-thrombin Binding To Nonpolymerized Fibrin-Sepharose: Evidence For An Anionic Binding Region.

L. Berliner, Y. Sugawara, J. Fenton
Published 1985 · Chemistry, Medicine

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In order to investigate ligand binding sites in alpha-thrombin that interact with nonpolymerized fibrin, fibrinogen was conjugated (with CNBr) to Sepharose 4B and converted to the nonpolymerized fibrin resin with alpha-thrombin. Human alpha-thrombin was bound to the resin at 22 degrees C and eluted with a linear NaCl gradient [50-300 mM in 50 mM tris(hydroxymethyl)aminomethane hydrochloride, pH 7.6] with midpeak elution occurring at an ionic strength that corresponds to 170 +/- 5 mM NaCl. Among various ligands examined, ATP and its analogues caused alpha-thrombin to elute with 125 mM or less salt. Apparent dissociation constants were estimated by the dependence of elution volume on ligand concentration. The most potent ligands for desorption from the column were anionic (e.g., adenine nucleotides), which also inhibit thrombin esterolytic/amidolytic and clotting activity [Conery, B. G., & Berliner, L. J. (1983) Biochemistry 22, 369-375]. The desorption series was at 10 mM concentrations: ATP = ADP greater than pyrophosphate greater than citrate greater than oxalate greater than PO4(3-). Contrastingly, serotonin and related apolar compounds did not cause dissociation of alpha-thrombin from the fibrin resin, even though several of these substances inhibit fibrinogen clotting and esterolytic/amidolytic activities of the enzyme. These data imply that independent sites for apolar and anionic binding in alpha-thrombin are required for converting fibrinogen into clottable fibrin and that alpha-thrombin-fibrin binding involves an anionic site.
This paper references
Human thrombins. Production, evaluation, and properties of alpha-thrombin.
J. Fenton (1977)
10.1016/0049-3848(73)90025-X
Adsorption of fibrinogen derivatives on insolubilized fibrinogen and fibrinmonomer
D. Heene (1973)
Human thrombins. Group IA and IIA salt-dependent properties of alpha-thrombin.
B. H. Landis (1981)
10.1021/BI00303A037
Proflavin binding within the fibrinopeptide groove adjacent to the catalytic site of human alpha-thrombin.
S. A. Sonder (1984)
Photoaffinity labeling of a single receptor for alpha-thrombin on mouse embryo cells.
D. Carney (1979)
Studies on the mechanism of thrombin. Interaction with fibrin.
M. Kamiński (1983)
10.1007/BF00222493
Structure-function relationships in human α- and γ-thrombins
L. Berliner (2004)
10.1038/275501A0
A two-step fibrinogen–fibrin transition in blood coagulation
B. Blombäck (1978)
10.1021/BI00640A015
Physical evidence for an apolar binding site near the catalytic center of human alpha-thrombin.
L. Berliner (1977)
10.1016/0049-3848(74)90023-1
Reduction of insolubilized fibrinogen.
F. Matthias (1974)
10.1111/J.1432-1033.1968.TB00473.X
Comparative studies on the inhibition of trypsin, plasmin, and thrombin by derivatives of benzylamine and benzamidine.
F. Markwardt (1968)
10.1021/BI00271A021
Binding subsites in human thrombins.
B. G. Conery (1983)
Selective immobilization of alpha-thrombin by surface-bound fibrin.
Wilner Gd (1981)
Steady state kinetic parameters for the thrombin-catalyzed conversion of human fibrinogen to fibrin.
Deborah L. Higgins (1983)
10.1016/0003-2697(85)90007-7
Gold labeling of thrombin and ultrastructural studies of thrombin-gold conjugate binding by fibrin.
C. Liu (1985)



This paper is referenced by
prothrombin molecules (Met-337-->Thr and Arg-388-->His) Prothrombin Himi: a compound heterozygote for two dysfunctional
Eriko Morishita (2011)
10.1016/J.THROMRES.2004.04.014
Standard measurement of clot-bound thrombin by using a chromogenic substrate for thrombin.
Sadia Meddahi (2004)
10.1111/J.1432-1033.1992.TB16916.X
The interaction of thrombin with fibrinogen. A structural basis for its specificity.
M. Stubbs (1992)
10.1021/BI00124A015
Conformational stability of a thrombin-binding peptide derived from the hirudin C-terminus.
F. Ni (1992)
10.1016/S0002-9149(98)00660-2
Direct thrombin inhibitors for treatment of arterial thrombosis: potential differences between bivalirudin and hirudin.
S. Bates (1998)
10.1074/JBC.M008249200
Unique Pathway of Thrombin-induced Platelet Aggregation Mediated by Glycoprotein Ib*
G. Soslau (2001)
10.1021/BI00421A027
Interaction of hirudin with thrombin: identification of a minimal binding domain of hirudin that inhibits clotting activity.
S. J. Mao (1988)
10.1111/j.1749-6632.1986.tb34577.x
Activation of Blood Coagulation Factor XIII a
L. Lorand (1986)
10.1021/BI00218A008
Characterization of the kinetic pathway for fibrin promotion of alpha-thrombin-catalyzed activation of plasma factor XIII.
M. Naski (1991)
10.1002/j.1552-4604.1994.tb03960.x
New Antithrombotic Strategies for Resistant Thrombotic Processes
L. Harker (1994)
10.1016/0014-2999(94)00464-1
Thrombin inhibitors and anti-coagulants on thrombin-induced embolisation in rabbit cranial vasculature.
J. Liu (1994)
10.1074/jbc.M600752200
The Structural Integrity of Anion Binding Exosite I of Thrombin Is Required and Sufficient for Timely Cleavage and Activation of Factor V and Factor VIII*
M. A. Bukys (2006)
10.1021/BI00418A066
Anion-binding exosite of human alpha-thrombin and fibrin(ogen) recognition.
J. Fenton (1988)
10.1111/J.1432-1033.1997.00550.X
Molecular cloning and expression of bothrojaracin, a potent thrombin inhibitor from snake venom.
V. Arocas (1997)
10.1021/BI00080A031
Thrombin-fibrinogen interaction: pH dependence and effects of the slow-->fast transition.
A. Mathur (1993)
10.1203/00006450-199605000-00014
Decreased Thrombin Activity of Fibrin Clots Prepared in Cord Plasma Compared with Adult Plasma
P. Patel (1996)
10.1016/0049-3848(90)90099-X
Zinc modulates thrombin adsorption to fibrin.
P. Hopmeier (1990)
10.1016/0049-3848(93)90002-6
A player of many parts: the spotlight falls on thrombin's structure.
M. Stubbs (1993)
10.1161/01.CIR.97.6.544
Thrombin binds to soluble fibrin degradation products where it is protected from inhibition by heparin-antithrombin but susceptible to inactivation by antithrombin-independent inhibitors.
J. Weitz (1998)
10.1111/J.1432-1033.1995.TB20785.X
Characterization of the binding of factor Xa to fibrinogen/fibrin derivatives and localization of the factor Xa binding site on fibrinogen.
M. Iino (1995)
10.1055/S-0038-1648875
Effects of the synthetic thrombin inhibitor argatroban on fibrin- or clot-incorporated thrombin: comparison with heparin and recombinant Hirudin.
C. N. Berry (1994)
10.1182/BLOOD.V71.3.556.556
Functional characterization of thrombin Salakta: an abnormal thrombin derived from a human prothrombin variant.
A. Bezeaud (1988)
Antiplatelet Versus Antithrombins
J. Chesebro (2005)
10.1111/j.1749-6632.1986.tb34570.x
Thrombin: Active‐Site Topography a
L. Berliner (1986)
10.1007/BF02171860
Hirudin and Hirulog: Advances in antithrombotic therapy
J. Maraganore (1994)
10.1074/jbc.M011324200
An Extensive Interaction Interface between Thrombin and Factor V Is Required for Factor V Activation*
Timothy Myles (2001)
10.1002/(SICI)1097-0134(19990101)34:1<29::AID-PROT4>3.0.CO;2-U
New general approach for determining the solution structure of a ligand bound weakly to a receptor: structure of a fibrinogen Aα‐like peptide bound to thrombin(S195A) obtained using NOE distance constraints and an ECEPP/3 flexible docking program
M. Maurer (1999)
10.1182/BLOOD.V90.5.1883
A novel degradation pathway of tissue factor pathway inhibitor: incorporation into fibrin clot and degradation by thrombin.
N. Ohkura (1997)
10.1016/0014-5793(91)80659-Q
Structure‐function relationships of hirulog peptide interactions with thrombin
P. Bourdon (1991)
10.1016/0049-3848(96)00111-9
Fibrinogen and fibrin--proteins with complex roles in hemostasis and thrombosis.
B. Blombäck (1996)
10.1006/ABIO.2000.4595
A fiber-optic microarray biosensor using aptamers as receptors.
M. Lee (2000)
10.1182/BLOOD-2005-01-0240
Studies on the basis for the properties of fibrin produced from fibrinogen-containing γ′ chains
K. Siebenlist (2005)
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