Haemophilia B: database of point mutations and short additions and deletions, fifth edition, 1994

Giannelli, F.; Green, P.M.; Sommer, Steve S.; Lillicrap, D.P.; Ludwig, M.; Schwaab, R.; Ph, Reitsma; Goossens, M.; Yoshioka, A; Brownlee, G. G.

doi:10.1093/nar/22.17.3534

Cited by 53 publications

(39 citation statements)

References 12 publications

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“…Naturally occurring 193 mutants are reported among coagulation factors, including Factor XI (13), Factor IX (14), and Factor VII as well (15). These enzymes were characterized biochemically and found to be altered in various aspects when compared with their Gly-193-possessing counterpart.…”

Section: From the Department Of Biochemistry Eötvös Loránd Universitmentioning

confidence: 99%

Thermodynamic Analysis Reveals Structural Rearrangement during the Acylation Step in Human Trypsin 4 on 4-Methylumbelliferyl 4-Guanidinobenzoate Substrate Analogue

Toth

Gombos

Simon

et al. 2006

Journal of Biological Chemistry

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Human trypsin 4 is an unconventional serine protease that possesses an arginine at position 193 in place of the highly conserved glycine. Although this single amino acid substitution does not affect steady-state activity on small synthetic substrates, it has dramatic effects on zymogen activation, interaction with canonical inhibitors, and substrate specificity toward macromolecular substrates. To study the effect of a non-glycine residue at position 193 on the mechanism of the individual enzymatic reaction steps, we expressed wild type human trypsin 4 and its R193G mutant. 4-Methylumbelliferyl 4-guanidinobenzoate has been chosen as a substrate analogue, where deacylation is rate-limiting, and transient kinetic methods were used to monitor the reactions. This experimental system allows for the separation of the individual reaction steps during substrate hydrolysis and the determination of their rate constants dependably. We suggest a refined model for the reaction mechanism, in which acylation is preceded by the reversible formation of the first tetrahedral intermediate. Furthermore, the thermodynamics of these steps were also investigated. The formation of the first tetrahedral intermediate is highly exothermic and accompanied by a large entropy decrease for the wild type enzyme, whereas the signs of the enthalpy and entropy changes are opposite and smaller for the R193G mutant. This difference in the energetic profiles indicates much more extended structural and/or dynamic rearrangements in the equilibrium step of the first tetrahedral intermediate formation in wild type human trypsin 4 than in the R193G mutant enzyme, which may contribute to the biological function of this protease.Trypsin is a prototype of the S1 serine protease family, the largest group of proteases. The residues indispensable for its enzymatic activity, His-57, , are structurally conserved and are referred to collectively as the catalytic triad (1). These residues are located at the active site of the enzyme. As the first step of the catalytic cycle, the enzyme binds the substrate forming the non-covalent Michaelis complex. The hydroxyl oxygen of the catalytic serine makes a nucleophilic attack on the electron-deficient carbonyl carbon of the scissile bond, and a covalent bond is formed. The attacked carbon atom becomes tetrahedrally coordinated; thus this state is called the first tetrahedral intermediate. In the next step, this species breaks down to yield the C-terminal (first) product and the acyl-enzyme. The acylation is followed by deacylation, a mechanistically similar action, in which the acyl-ester bond is attacked by a water molecule that has previously been activated by 3).During the hydrolysis of the scissile bond, tetrahedral intermediates are developed that are stabilized via hydrogen bonding interactions, in which the amido groups of residues Gly-193 and Ser-195 act as H-donors and the developing oxyanion intermediate is the acceptor (4, 5). Despite glycine being the far most convenient amino acid at position 193 for optimal...

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Section: From the Department Of Biochemistry Eötvös Loránd Universitmentioning

confidence: 99%

Thermodynamic Analysis Reveals Structural Rearrangement during the Acylation Step in Human Trypsin 4 on 4-Methylumbelliferyl 4-Guanidinobenzoate Substrate Analogue

Toth

Gombos

Simon

et al. 2006

Journal of Biological Chemistry

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“…Factor IX (F.IX) 1 is a 57-kDa zymogen of a serine protease that participates in blood coagulation. Activated F.IX (factor IXa), with its nonenzymatic cofactor, factor VIIIa, activates factor X to factor Xa, which converts prothrombin to thrombin in the penultimate step of the coagulation cascade.…”

mentioning

confidence: 99%

“…Activated F.IX (factor IXa), with its nonenzymatic cofactor, factor VIIIa, activates factor X to factor Xa, which converts prothrombin to thrombin in the penultimate step of the coagulation cascade. Mutations in F.IX result in the bleeding disorder hemophilia B (1).…”

mentioning

confidence: 99%

Human Factor IX Binds to Specific Sites on the Collagenous Domain of Collagen IV

Wolberg¹,

Stafford²,

Erie³

1997

Journal of Biological Chemistry

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The primary region of factor IX that mediates binding to bovine aortic endothelial cells resides in residues 3-11 of the N-terminal region known as the Gla domain. Recently, it was proposed that the observed binding to endothelial cells is actually a measure of the interaction between factor IX and collagen IV (Cheung, W. F., van den Born, J., Kuhn, K., Kjellen, L., Hudson, B. G., and Stafford, D. W. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 11068 -11073). To confirm that factor IX binds to collagen IV and to examine the specificity of this interaction, we used scanning force microscopy to examine factor IX binding to collagen IV. We imaged collagen IV in the presence and the absence of factor IX and observed specific interactions between factor IX and collagen IV. Our results demonstrate that factor IX binds to collagen IV at specific sites in the collagenous domain ϳ98 and ϳ50 nm from the C-terminal pepsin-cleaved end.

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“…Alanine** is the second residue downstream of the aspartic acid, which forms part of the catalytic triad 242, Ser-344; it is also highly conserved in the vitamin K-dependent coagulation factors of both human and bovine origin. A mutation analogous to FVII Malta I1 has been detected in four unrelated cases with hemophilia B (fifth edition of the hemophilia B database) (Giannelli et al, 1994). In these patients A271V was associated with markedly decreased F1X:C and FIX:Ag (averagks of reported F K C and FIX:Ag for these cases are 12% and 4%, respectively).…”

Section: S191mentioning

confidence: 98%

“…A mutation at the corresponding position has been detected in the human FIX gene (P131L; fifth edition of the hemophilia B database) (Giannelli et al, 1994). However, no phenotype data are available for this mutation.…”

Section: S191mentioning

confidence: 99%

Two new missense mutations (P134T and A244V) in the coagulation factor VII gene

et al. 1998

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Haemophilia B: database of point mutations and short additions and deletions, fifth edition, 1994

Cited by 53 publications

References 12 publications

Thermodynamic Analysis Reveals Structural Rearrangement during the Acylation Step in Human Trypsin 4 on 4-Methylumbelliferyl 4-Guanidinobenzoate Substrate Analogue

Thermodynamic Analysis Reveals Structural Rearrangement during the Acylation Step in Human Trypsin 4 on 4-Methylumbelliferyl 4-Guanidinobenzoate Substrate Analogue

Human Factor IX Binds to Specific Sites on the Collagenous Domain of Collagen IV

Two new missense mutations (P134T and A244V) in the coagulation factor VII gene

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