Two Naturally Occurring Mutations on FVII Gene (S363I-W364C) Altering Intrinsic Catalytic Activity

Flora Peyvandi; Raimondo De Cristofaro; Sepideh Akhavan; Josephine A. Carew; Raffaele Landolfi; Kenneth A. Bauer; Pier Mannuccio Mannucci

doi:10.1055/s-0037-1613297

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2002; 88(05): 750-755
DOI: 10.1055/s-0037-1613297

Review Article

Schattauer GmbH

Two Naturally Occurring Mutations on FVII Gene (S363I-W364C) Altering Intrinsic Catalytic Activity

Flora Peyvandi

¹Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, Fondazione Luigi Villa, IRCCS Maggiore Hospital and University of Milan, Italy

,

Raimondo De Cristofaro

²Hemostasis Research Center, Catholic University School of Medicine, Rome, Italy

,

Sepideh Akhavan

¹Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, Fondazione Luigi Villa, IRCCS Maggiore Hospital and University of Milan, Italy

,

Josephine A. Carew

³Hematology Section, Department of Medicine, VA Boston Healthcare System, Harvard Medical School, Boston, MA, USA

,

Raffaele Landolfi

²Hemostasis Research Center, Catholic University School of Medicine, Rome, Italy

,

Kenneth A. Bauer

³Hematology Section, Department of Medicine, VA Boston Healthcare System, Harvard Medical School, Boston, MA, USA

,

Pier Mannuccio Mannucci

¹Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, Fondazione Luigi Villa, IRCCS Maggiore Hospital and University of Milan, Italy

› Institutsangaben

Abstract

Summary

Factor VII (FVII) requires the cleavage of an internal peptide bond and the association with tissue factor (TF) to attain its fully active FVIIa conformation. This event alone leaves FVIIa in a zymogen-like state of relatively low specific activity. The TF-induced allosteric enhancement of FVIIa’s activity contributes to the procoagulant activity of the complex. We have characterized two naturally occurring mutations (S363I W364C) on FVII gene. Both homozygous patients for each mutation have a normal FVII:Ag level associated to an undetectable FVII coagulant activity. The patient carrying the allele 364C had a more severe hemorrhagic diathesis than the S363I mutant. To understand the mechanism of these deficiency, in vitro expression analysis with further biochemical characterization of recombinant proteins of both mutants FVII-363I, FVII-364C and wild type (WTFVII) FVII constructs were done. The results recapitulated the patients’ plasma data with normal Ag level and no detectable coagulant activity. The D-F-Pip-R-pNA and CH3SO2-D-CHA-A-But-R chromogenic substrates were used to evaluate the amidolytic activity of WT and mutant FVII in presence and absence of recombinant tissue factor (rTF). Binding of FVII to rTF by a solid phase binding assay was done using recombinant human rTF. The results of amidolytic assays showed that rTF enhances 28 fold the value of the specificity of constant (kcat/Km) in WT but no activity was detectable in either mutant constructs under any condition. The equilibrium dissociation constant of rTF-FVIIa interaction showed Kd equal to 4.4 ± 0.2nM, 4.9 ± 0.5nM and 6 ± 0.9 of WT, 363I and 364C FVII forms, respectively. The Kd values of the non activated forms were equal to 24.7 ± 3.3, 24.4 ± 3.1 and 20.6 ± 4nM, respectively. These data demonstrate that, compared to the WT form, FVII-363I and FVII-364C have no significant affinity change for TF and that the detrimental effect of these two mutations is attributable to the loss of an efficient catalytic machinery in the FVII molecule causing a severe deficiency of coagulant activities.

Keywords

Factor VII - extrinsic coagulation - tissue factor

Volltext

Referenzen

References
1 Peyvandi F, Mannucci PM, Asti D, Abdoullahi M, Di Rocco N, Sharifian R. Clinical manifestation in 28 Italian and Iranian patients with severe factor VII deficiency. Haemophilia 1997; 03: 242-6.
2 Cooper DN, Millar DS, Wacey A, Banner DW, Tuddenham EGD. Inherited factor VII deficiency: molecular genetics and pathophysiology. Thromb Haemost 1997; 78: 151-60.
3 Peyvandi F, Jenkins PV, Mannucci PM, Billio A, Zeinali S, Perkins SJ, Perry DJ. Molecular characterisation and three-dimensional structural analysis of mutations in 21 unrelated families with inherited Factor VII deficiency. Thromb Haemost 2000; 84: 250-7.
4 Leonard B, Chen Q, Blajchman M, Ofosu F, Sridhara S, Yang D, Clarke B. Factor VII deficiency caused by a structural variant N57D of the first epidermal growth factor domain. Blood 1998; 91: 142-8.
5 O’Brien DP, Kembal-Cook G, Hutchinson AM, Martin DMA, Jhonson DJD, Byfield PGH, Takamiya O, Tuddenham EGD, Mc Vey JH. Surface plasmon resonance studies of the interaction between factor VII and tissue factor. Demonstration of defective tissue factor binding in a variant FVII molecule (FVII-R79Q). Biochemistry 1994; 33: 14162-9.
6 Kazama Y, Foster DC, Kisiel W. Evidence that an Arg79→Gln substitution in human factor VII is not associated with a reduction in coagulant activity. Blood Coagul Fibrinolysis 1992; 03: 697-702.
7 Sridhara S, Clarke BJ, Blajchman MA. Arginine-79 in the first epidermal growth factor domain of factor VII is essential for the interaction with tissue factor. Blood Coagul Fibrinolysis 1993; 04: 505-6.
8 Chaing S, Clarke B, Sridhara S, Chu K, Friedman P, Vandusen W, Roberts HR, Blajchman M, Moroe DM, High KA. Severe factor VII deficiency caused by mutations abolishing the cleavage site for activation and altering binding to tissue factor. Blood 1994; 83: 3524-35.
9 Kemball-Cook G, Jhonson JD, Takamiya O, Banner DW, McVey J, Tuddenham EGD. Coagulation factor VII Glu100→Arg. J Biol Chem 1998; 273: 8516-21.
10 Hanault M, Arbini AA, Carew JA, Peyvandi F, Bauer KA. Characterization of two naturally occurring mutations in the second epidermal growth factorlike domain of factor VII. Blood 1999; 93: 1237-44.
11 Wilgoose P, Berkner KL, Kisiel W. Synthesis, purification, and characterization of an Arg152→Glu site-directed mutant of recombinant human blood clotting factor VII. Biochemistry 1990; 29: 3413-20.
12 Matsushita T, Kojima T, Emi N, Takahashi I, Saito H. Impaired human tissue factor mediated activity in blood clotting factor VII Nagoya (Arg304Trp). Evidence that a region in the catalytic domain of factor VII is important for the association with tissue factor. J Biol Chem 1994; 269: 7355-63.
13 Bharadwaj D, Ino M, Kontoyianni M, Smith KJ, Foster DC, Kisiel W. Factor VII central. A novel mutation in the catalytic domain that reduces tissue factor binding, impaires activation by factor Xa, and abolishes amidolytic and coagulant activity. J Biol Chem 1996; 271: 30685-91.
14 Coppola R, Tombesi S, Valentini F, Alborali S, Albertini A, Mannucci PM. Enzyme-linked immunosorbent assay of human factor VII based upon a monoclonal antibody that recognizes the native conformation of the protein. Thromb Res 1992; 68: 283-93.
15 Banner DW, D’Arcy A, Chene C. et al. The crystal structure of the complex of blood coagulation factor VIIa with soluble tissue factor. Nature 1996; 380: 41-6.
16 Arbini AA, Mannucci PM, Bauer KA. A Thr359Met mutation in factor VII of a patient with a hereditary deficiency causes defective secretion of the molecule. Blood 1996; 87: 5085-94.
17 Kaufman RJ, Davies MV, Waslei LC, Michnick D. Improved vectors for stable expression of foreign genes in mammalian cells by use of the untranslated leader sequence from EMC virus. Nucleic Acids Res 1991; 19: 4485-90.
18 Peyvandi F, Carew JA, Perry DJ, Hanault M, Khanduri U, Perkins SJ, Mannucci PM, Bauer KA. Abnormal secretion and function of recombinant human factor VII as the result of modification to a calcium binding site caused by a 15 bp insertion in the factor VII gene. Blood 2001; 96: 960-5.
19 Urlaub G, Chasin LA. Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity. Proc Natl Acad Sci Usa 1980; 77: 4216-20.
20 Broze GJ, Hickman S, Miletich JP. Monoclonal Anti-human Factor VII antibodies. Detection in plasma of a second protein antigenically and genetically related to FVII. J Clin Invest 1985; 76: 937-46.
21 Bajaj SP, Rapaport SI, Brown SF. Isolation and characterization of human factor VII. Activation of factor VII by factor Xa. J Biol Chem 1981; 256: 253-9.
22 Higashi S, Matsumoto N, Iwanaga S. Molecular mechanism of tissue factor-mediated acceleration of factor VIIa activity. J Biol Chem 1996; 271 (43) 26569-74.
23 Silverberg SA, Nemerson Y, Zur M. Kinetics of the activation of bovine coagulation factor X by components of the extrinsic pathway. Kinetic behavior of two-chain factor VII in the presence and absence of tissue factor. J Biol Chem 1977; 252: 8481-8.
24 Ashton AW, Boehm MK, Johnson DJD, Kemball-Cook G, Perkins SJ. The solution structure of human coagulation factor VIIa in its complex with tissue factor: A study of a heterodimeric receptor-ligand complex by X-ray and neutron scattering and computational modelling. Biochemistry 1998; 37: 8208-17.
25 Perkins SJ, Smith KF. Identity of the putative serine-proteinase fold in proteins of the complement system with nine relevant crystal structures. Biochem J 1993; 295: 109-14.
26 Derewenda ZS, Derewenda U, Kobos P. (His)C_ε-H…O = C< hydrogen bond in the cative sites of serine hydrolases. J Mol Biol 1994; 241: 83-93.
27 Engh RA, Brandstetter H, Sucher G, Eichinger A, Baumann U, Bode W, Huber R, Poll T, Rudolph R, von der Saal W. Enzyme flexibility, solvent and “weak” interactions characterize thrombin-ligand interactions: implications for drug design. Structure 1996; 04: 1353-62.
28 Bode W, Turk D, Karshikov A. The refined 1.9 Å crystal structure of D PheProArgchloromethylketone inhibited human α-thrombin. Structure analysis, overall structure, electrostatic properties, detailed active site geometry, structure-function relationships. Protein Sci 1992; 01: 426-71.
29 Kemball-Cook G, Johnson DJD, Tuddenham EGD, Harlos K. Crystal structure of active site-inhibited human coagulation factor VIIa (des-Gla). J Struct Biol 1991; 27: 213-23.