The mechanism of the fibrinogen-thrombin reaction

Hogg, Desmond H.; Blombäck, Birger

doi:10.1016/0049-3848(78)90051-8

Cited by 54 publications

(40 citation statements)

References 17 publications

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“…They proposed that the region 30-44 contains amino acid residues that are important for the interaction with thrombin [13]. The kinetic parameters obtained in this paper for Aa(1-44) ( Table 2) are in agreement with this hypothesis.…”

Section: Discussionsupporting

confidence: 80%

“…The close similarity of the kinetic parameters for Aa(1-23) and Aa(1-29) and the absence of an effect of TM on the hydrolysis of both substrates is in agreement with the proposal by Hogg and Blomback [13] that the region 23-33 of the Aa chain does not contribute significantly to the enzymesubstrate interaction. They proposed that the region 30-44 contains amino acid residues that are important for the interaction with thrombin [13].…”

Section: Discussionsupporting

confidence: 78%

“…The highly conserved nature of Phe-8 (position P9) during the evolution of the fibrinogen molecule has suggested the importance of this residue in the enzyme substrate interaction [35]. Experimental confirmation of this hypothesis was obtained from the comparison of the kinetic parameters for the cleavage of the synthetic peptides Aa (8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20), Aa (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) and Aa(10-20) [18]. Tripeptidyl pnitroanilide substrates having a D-Phe residue at the P3 position were synthesized based on this rationale.…”

Section: Discussionmentioning

confidence: 51%

“…The mixture was fractionated by reverse-phase chromatography on a CI8 column under conditions similar to those used in the purification of the fragments (described in the miniprint section). Detectable amounts of Aa (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19) and Aa were not found (a small amount of cleavage at Arg-16 was detected, due to the presence of approximately 1% of free thrombin). Moreover, the peak containing the substrate was shown to display the N-terminal sequence of Aa(l-44), i.e.…”

Section: The Effect Of Tm On the Cleavage Of The Various Forms Offibrmentioning

confidence: 99%

“…It was found that the N-terminal 51 amino acid residues of the Aa chain of fibrinogen contain all the information for the specific interaction between thrombin and the Aa chain of fibrinogen [13,151. By using smaller fragments, regions within this sequence, and even single amino acid residues that play a role in the enzyme-substrate interaction, could be identified In order to examine the question of whether TM influences the specific interactions between thrombin and fibrinogen we studied the effect of TM from rabbit lung on the release of [15 -201. fibrinopeptides by human thrombin from fragments of the human fibrinogen molecule.…”

mentioning

confidence: 99%

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The effect of thrombomodulin on the cleavage of fibrinogen and fibrinogen fragments by thrombin

Hofsteenge

Stone

1987

Eur J Biochem

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Thrombomodulin acts as a linear competitive inhibitor of thrombin with respect to the substrate fibrinogen. In the present study the effect of thrombomodulin on the activity of thrombin with fragments of the Aa and B/? chain of fibrinogen has been examined. The cleavage of fibrinopeptide A from the N-terminal disulphide knot, fragment 1 -44 and fragment 1 -51 of the Aa chain was inhibited by thrombomodulin. The average value for the inhibition constant obtained with these substrates was 0.83 0.09 nM, which was in good agreement with the values obtained previously for the inhibition of thrombin by thrombomodulin with native fibrinogen as the substrate [Hofsteenge, J., Taguchi In addition, we determined the second-order rate constants of cleavage of these substrates using human thrombin. Fragments of the Aa chain whose cleavage was inhibited by thrombomodulin were found to have values for k,,,/K,,, that were within one order of magnitude of that for the native fibrinogen, whereas those for Aa chain fragments whose cleavage was not inhibited by thrombomodulin were found to be more than two orders of magnitudes lower.From these results we conclude that only a relatively small portion of the A a chain of the fibrinogen molecule is responsible for the specific binding to thrombin that is affected by thrombomodulin. Moreover, residues 30 -44 of the Aa chain play an important role in this thrombin-fibrinogen interaction.The interaction of thrombin with thrombomodulin (TM) is one possible mechanism for the regulation of the level of active thrombin in the blood coagulation system [l, 21. Thronibomodulin is an endothelial cell-surface protein that forms a complex with thrombin, with a dissociation constant of approximately 0.7 nM [3 -51. The protein has been purified from rabbit and bovine lung, and human placenta and has an apparent relative molecular mass of 64000 -105000, depending on the source [4, 6, 71. Thrombin complexed to TM has a changed specificity towards macromolecular substrates. In contrast to free thrombin, the complex cannot cleave fibrinogen and factor V nor can it activate platelets, but it can efficiently activate protein C [l, 3, 8 -101. Activated protein C subsequently can inactivate factor V,, a factor indispensable for the conversion of prothrombin into thrombin by factor X,. Thus the binding of thrombin to thrombomodulin results in inhibition of both the formation and the procoagulant activities of this enzyme. The mechanism by which the change in specificity occurs is only partially understood. TM from bovine and rabbit lung are linear competitive inhibitors of thrombin with respect to fibrinogen [4, 51. In contrast, the hydrolysis of tripeptidyl p-nitroanilide substrates by thrombin is not inhibited by the binding to TM 14, 5, 81. Similarly, TM competes for the binding of the polypeptide inhibitor hirudin (65 residues) to thrombin, but it does not significantly alter the rate of inactivation of thrombin by tripeptidyl chloromethane inhibitors [4, 51. These findings show that TM binds to, or al...

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Section: Discussionsupporting

confidence: 80%

Section: Discussionsupporting

confidence: 78%

Section: Discussionmentioning

confidence: 51%

Section: The Effect Of Tm On the Cleavage Of The Various Forms Offibrmentioning

confidence: 99%

mentioning

confidence: 99%

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The effect of thrombomodulin on the cleavage of fibrinogen and fibrinogen fragments by thrombin

Hofsteenge

Stone

1987

Eur J Biochem

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From natural to synthetic multisite thrombin inhibitors

et al. 1999

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Blood Coagulation and Fibrinolysis

Colman

Budzynski

1985

Comprehensive Physiology

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The sections in this article are: Congenital Deficiencies in Initiation of Blood Coagulation Factor XII Plasma Prekallikrein Plasma Kininogens Interaction of Contact‐System Proteins Factor XI Factor IX Factor VIII Relationship of Intrinsic and Extrinsic Blood Coagulation Factor X Factor VII Thromboplastin Regulation of Prothrombin Activation Prothrombin Factor V Coagulant Phospholipid Surfaces Calcium Ions Thrombin Formation Thrombin Specificity Regulation of Thrombin Activity Heparin and Antithrombin III Interaction of Thrombin With Fibrin Nonmammalian Inhibitors of Thrombin Thrombinlike Enzymes in Snake Venoms Electron Microscopy of Fibrinogen Fibrinogen Structure Model of Fibrinogen Molecule Conversion of Fibrinogen to Fibrin Clot Polymerization Sites Cross‐Linking of Fibrin Interaction of Fibrinogen with Plasma Proteins Interaction of Fibrinogen with Cells Degradation by Proteases other than Plasmin Degradation of Fibrinogen and Fibrin by Plasmin Role of Fibrin in Clot Formation and Lysis Plasminogen and Plasmin Plasminogen Activators Inhibitors of Plasminogen and Plasmin Physiological Fibrinolysis

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The mechanism of the fibrinogen-thrombin reaction

Cited by 54 publications

References 17 publications

The effect of thrombomodulin on the cleavage of fibrinogen and fibrinogen fragments by thrombin

The effect of thrombomodulin on the cleavage of fibrinogen and fibrinogen fragments by thrombin

From natural to synthetic multisite thrombin inhibitors

Blood Coagulation and Fibrinolysis

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