Factor VIII Arg2304 → His Binds to Phosphatidylserine and Is a Functional Cofactor in the Factor X-ase Complex

Deborah A. Lewis; Karen D. Moore; Thomas L. Ortel

doi:10.1055/s-0037-1615697

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2001; 85(02): 260-264
DOI: 10.1055/s-0037-1615697

Review Article

Schattauer GmbH

Factor VIII Arg₂₃₀₄ → His Binds to Phosphatidylserine and Is a Functional Cofactor in the Factor X-ase Complex

Autoren

Deborah A. Lewis

¹Department of Medicine, Division of Hematology, and the Department of Pathology, Duke University Medical Center, Durham, NC, USA
Karen D. Moore

¹Department of Medicine, Division of Hematology, and the Department of Pathology, Duke University Medical Center, Durham, NC, USA
Thomas L. Ortel

¹Department of Medicine, Division of Hematology, and the Department of Pathology, Duke University Medical Center, Durham, NC, USA

Abstract

Summary

Four factor VIII light chain constructs containing hemophilia A mutations at R2304 and R2307 were prepared and expressed in mammalian cells. These mutations are located in a putative phosphatidylserine binding site identified by peptide studies (spanning amino acids 2303-2332). The levels of all four mutants in conditioned medium were significantly less than wild type by immunoprecipitation and ELISA. R2304H and wild type factor VIII light chains were concentrated by cation exchange chromatography from medium. R2304H and wild type factor VIII light chains bound immobilized phosphatidylserine similarly. The reconstituted cofactor activity of R2304H factor VIII light chain was slightly greater than wild type factor VIII light chain. These results are consistent with the recently reported crystal structure of factor VIII C2 domain that suggests R2304H is not directly involved in phospholipid binding. The observed clinical phenotype is probably due to decreased circulating levels of a functional protein.

Keywords

Factor VIII - factor VIII light chain - hemophilia - phospholipid binding

Volltext

Referenzen

References
1 Lenting PJ, van Mourik JA, Mertens K. The life cycle of coagulation factor VIII in view of its structure and function. Blood 1998; 92: 3983-96.
2 Fay PJ, Haidaris PJ, Smudzin TM. Human factor VIIIa subunit structure. Reconstitution of factor VIII_a from the isolated A1/A3-C1-C2 dimer and A2 subunit. J Biol Chem 1991; 266: 8957-62.
3 Kane WH, Davie EW. Blood coagulation factors V and VIII: structural and functional similarities and their relationship to hemorrhagic and thrombotic disorders. Blood 1988; 71: 539-55.
4 Arai M, Scandella D, Hoyer LW. Molecular basis of factor VIII inhibition by human antibodies. Antibodies that bind to the factor VIII light chain prevent the interaction of factor VIII with phospholipid. J Clin Invest 1989; 83: 1978-84.
5 Nesheim ME, Pittman DD, Wang JH, Slonosky D, Giles AR, Kaufman RJ. The binding of ³⁵S-labeled recombinant factor VIII to activated and unactivated human platelets. J Biol Chem 1988; 263: 16467-70.
6 Ortel TL, Devore-Carter D, Quinn-Allen MA, Kane WH. Deletion analysis of recombinant human factor V. Evidence for a phosphatidylserine binding site in the second C-type domain. J Biol Chem 1992; 267: 4189-98.
7 Tracy PB, Mann KG. Prothrombinase complex assembly on the platelet surface is mediated through the 74,000-dalton component of factor Va. Proc Natl Acad Sci USA 1983; 80: 2380-4.
8 Foster PA, Fulcher CA, Houghten RA, Zimmerman TS. Synthetic factor VIII peptides with amino acid sequences contained within the C2 domain of factor VIII inhibit factor VIII binding to phosphatidylserine. Blood 1990; 75: 1999-2004.
9 Scandella D, Gilbert GE, Shima M, Nakai H, Eagleson C, Felch M, Prescott R, Rajalakshmi KJ, Hoyer LW, Saenko E. Some factor VIII inhibitor antibodies recognize a common epitope corresponding to C2 domain amino acids 2248 through 2312, which overlap a phospholipid-binding site. Blood 1995; 86: 1811-9.
10 Macedo-Ribeiro S, Bode W, Huber R, Quinn-Allen MA, Kim SW, Ortel TL, Bourenkov GP, Bartunik HD, Stubbs MT, Kane WH, Fuentes-Prior P. Crystal structures of the membrane-binding C2 domain of human coagulation factor V. Nature 1999; 402: 434-9.
11 Pratt KP, Shen BW, Takeshima K, Davie EW, Fujikawa K, Stoddard BL. Structure of the C2 domain of human factor VIII at 1.5 A resolution. Nature 1999; 402: 439-42.
12 Hoyer LW. Hemophilia A. N Engl J Med 1994; 330: 38-47.
13 Kemball-Cook G, Tuddenham EG, Wacey AI. The factor VIII structure and mutation resource site: HAMSTeRS version 4. Nucleic Acids Res 1998; 26: 216-9.
14 Pipe SW, Kaufman RJ. Factor VIII C2 domain missense mutations exhibit defective trafficking of biologically functional proteins. J Biol Chem 1996; 271: 25671-6.
15 Saenko EL, Scandella D. The acidic region of the factor VIII light chain and the C2 domain together form the high affinity binding site for von Wille-brand factor. J Biol Chem 1997; 272: 18007-14.
16 Nordfang O, Ezban M, Favaloro JM, Dahl HH, Hansen JJ. Specificity of monoclonal antibodies to factor VIII:C. Thromb Haemost 1985; 54: 586-90.
17 Ortel TL, Moore KD, Ezban M, Kane WH. Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII. Thromb Haemost 1996; 75: 36-44.
18 Kane WH, Devore-Carter D, Ortel TL. Expression and characterization of recombinant human factor V and a mutant lacking a major portion of the connecting region. Biochemistry 1990; 29: 6762-8.
19 Saenko EL, Scandella D. A mechanism for inhibition of factor VIII binding to phospholipid by von Willebrand factor. J Biol Chem 1995; 270: 13826-33.
20 Scandella D, Mattingly M, Prescott R. A recombinant factor VIII A2 domain polypeptide quantitatively neutralizes human inhibitor antibodies that bind to A2. Blood 1993; 82: 1767-75.
21 Fay PJ. Reconstitution of human factor VIII from isolated subunits. Arch Biochem Biophys 1988; 262: 525-31.
22 Nordfang O, Ezban M. Generation of active coagulation factor VIII from isolated subunits. J Biol Chem 1988; 263: 1115-8.
23 Prowse C, Hornsey V, McKay G, Waterston Y. Room temperature, microtray chromogenic assay of factor VIII:C. Vox Sang 1986; 50: 21-5.
24 Gale AJ, Pellequer J-L, Getzoff ED, Griffin JH. Structural basis for hemophilia A caused by mutations in the C domains of blood coagulation factor VIII. Thromb Haemost 2000; 83: 78-85.
25 Cameron C, Notley C, Hoyle S, McGlynn L, Hough C, Kamisue S, Giles A, Lillicrap D. The canine factor VIII cDNA and 5′ flanking sequence. Thromb Haemost 1998; 79: 317-22.
26 Kim SW, Quinn-Allen MA, Camp JT, Macedo-Ribeiro S, Fuentes-Prior P, Bode W, Kane WH. Identification of functionally important amino acid residues within the C2 domain of human factor V using alanine scanning mutagenesis. Biochemistry 2000; 39: 1951-8.
27 Takeshima K, Fujikawa K. The phospholipid binding property of the C2 domain of human factor VIII. Blood 1999; 82 Supplement 234-5.
28 Voorberg J, de Laaf RTM, Koster PM, van Mourik JA. Intracellular retention of a factor VIII protein with an Arg²³⁰⁷→ Gln mutation as a cause of haemophilia A. Biochem J 1996; 318: 931-7.