Phospholipid vesicles interfere with the binding of antibody fragments to the light chain of factor VIII

Summary

Factor VIII binds to phospholipid membranes through the C2 domain (S2173-Y2332). Residues M2199, F2200, L2251, L2252, V2223, W2313 and V2314 at the tips of β-hairpins and loops are thought to contribute to phospholipid membrane binding. Similarly, residues in the C2 domain of the homologous protein factor V form a phospholipid binding site, but residues in the A3 and C1 domains are also thought to contribute to membrane binding. Phage display technology was previously used to isolate factor VIII light chain specific single-chain variable domain fragments (scFv) from patients with factor VIII inhibitors. Phospholipid vesicles inhibited the binding of factor VIII to scFvsWR1 and WR16 (epitope: E2181-M2199) with half saturation values of 23 and 47 μM respectively. The single point mutant F2200A factor VIII light chain bound to WR1 and WR16 with a much lower affinity than wild type protein suggesting that residue F2200 is also included in the epitopes of these scFvs. Binding of factor VIII to C2-specific scFvsWR13 and EL14 (epitope: K2207-M2321) was not inhibited by phospholipid vesicles. Consistent with this, F2200A factor VIII light chain bound to these scFvs with the same affinity as the wild type protein. However, phospholipid vesicles also inhibited the binding of factor VIII to the A3-C1-specific scFvs KM36 (epitope: Q1778-D1840) and KM38 (epitope: S1690-N1777 and/or V1841-N2172) with half saturation values of 84 and 165 μM, respectively, suggesting that the A3 and/or C1 domains may contribute to membrane binding of the cofactor.

Keywords

Factor VIII - phospholipids - antibody

• References

• 1 Sadler JE, Davie EW, Hemophilia A. Hemophilia B and von Willebrand's disease. In: Stamatoyannopoulos G, Majerus PW, Perlmutter RM, Varmus H, editors. The molecular basis of blood diseases. Philadelphia: W. B. Saunders 2001; 680-718.
• 2 Foster PA, Fulcher CA, Houghten RA. et al. 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.
• 3 Nogami K, Shima M, Hosokawa K. et al. Factor VIII C2 domain contains the thrombin-binding site responsible for thrombin-catalyzed cleavage at Arg 1689. J Biol Chem 2000; 275: 25774-80.
• 4 Lenting PJ, van de Loo J-WHP, Donath M-JSH. et al. The sequence Glu1811-Lys1818of humanblood coagulation factor VIII comprises a binding site for activated factor IX. J Biol Chem 1996; 271: 1935-40.
• 5 Kalafatis M, Jenny RJ, Mann KG. Identification and characterization of a phospholipid-binding site of bovine factor Va. J Biol Chem 1990; 265: 21580-9.
• 6 Kalafatis M, Rand MD, Mann KG. Factor Va-membrane interaction ismediated by two regions located on the light chain of the cofactor. Biochemistry 1994; 33: 486-93.
• 7 Saleh M, Peng W, Quinn-Allen MA. et al. The factor V C1 domain is involved in membrane binding: identification of functionally important amino acid residueswithin the C1 domain of factor Vusing alanine scanning mutagenesis. Thromb Haemost 2004; 91: 16-27.
• 8 Kim SW, Quinn-Allen MA, Camp JT. et al. Identification of functionally important amino acid residues within the C2 Domain of human factor Vusing alanine scanning mutagenesis. Biochemistry 2000; 39: 1951-8.
• 9 Pratt KP, Shen BW, Takeshima K. et al. Structure of the C2 domain of human factor VIII at 1.5Aresolution. Nature 1999; 402: 439-42.
• 10 Macedo-Ribeiro S, Bode W, Huber R. et al. Crystal structures of the membrane-binding C2 domain of human coagulation factor V. Nature 1999; 402: 434-9.
• 11 Stoilova-McPhie S, Villoutreix BO, Mertens K. et al. 3-Dimensional structure of membrane-bound coagulation factor VIII: modeling of the factor VIII heterodimer within a 3-dimensional density map derived by electron crystallography. Blood 2002; 99: 1215-23.
• 12 Scandella D. Properties of anti-factor VIII inhibitor antibodies in hemophilia A patients. Semin Thromb Hemost 2000; 26: 137-42.
• 13 Fijnvandraat K, Celie PHN, Turenhout EAM. et al. Ahuman alloantibody interferes with binding of factor IXa to the factor VIII light chain. Blood 1998; 91: 2347-352.
• 14 Arai M, Scandella D, Hoyer W. 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.
• 15 Jacquemin MG, Desqueper BG, Benhida A. et al. Mechanism and kinetics of factor VIII inactivation: study with an IgG4 monoclonal antibody derived from a hemophilia A patient with inhibitor. Blood 1998; 92: 496-506.
• 16 Spiegel PC, Jacquemin M, Saint-Remy J-MR. et al. Structure of a factor VIII C2 domain-immunoglobulin G4k Fab complex: identification of an inhibitory anti body epitope on the surface of factor VIII. Blood 2001; 98: 13-19.
• 17 van den Brink EN, Turenhout EA, Bovenschen N. et al. Multiple VH genes are used to assemble human antibodies directed toward the A3-C1 domains of factor VIII. Blood 2001; 97: 966-72.
• 18 van den Brink EN, Bril WS, Turenhout EAM. et al. Two classes of germline genes both derived from the V(H)1 family direct the formation of human antibodies that recognize distinct antigenic sites in the C2 domain of factor VIII. Blood 2002; 99: 2828-34.
• 19 van den Brink EN, Turenhout EAM, Davies J. et al. Human antibodies with specificity for the C2 domain of factor VIII are derived from VH1 germline genes. Blood 2000; 95: 558-63.
• 20 Kim SW, Ortel TL, Quinn-Allen MA. et al. Partial glycosylation at asparagine-2181 of the second C-type domain of human factor V modulates assembly of the prothrombinase complex. Biochemistry 1999; 38: 11448-54.
• 21 Gomori G. A modification of the colorimetric phosphorus determination for use with the photoelectric colorimeter. J Lab Clin Med 1942; 27: 955-60.
• 22 Lewis DA, Pound ML, Ortel TL. Contributions of Asn²¹⁹⁸, Met²¹⁹⁹, and Phe²²⁰⁰ in the factor VIII C2 domain to cofactor activity, phospholipid-binding, and von Willebrand factor-binding. Thromb Haemost 2002; 89: 795-802.
• 23 Lewis DA, Moore KD, Ortel TL. Binding of factor VIII inhibitors to discrete regions of the factor VIII C2 domain disrupt phospholipid binding. Blood Coag Fibrinol 2003; 14: 361-8.
• 24 Mertens K, Donath MJSH, Van Leen RW. et al. Biological activity of recombinant factor VIII variants lacking the central B-domain and the heavy-chain sequence Lys⁷¹³-Arg⁷⁴⁰: Discordant in vitro and in vivo activity. Br J Haematol 1993; 85: 133-42.
• 25 Tao BY, Lee KCP, In Griffin HG, Griffin AM. editors PCRTechnology Current Inovations. Boca Raton: CRC Press 1994; 71-2.
• 26 Villard S, Piquer D, Raut S. et al. Low molecular weight peptides restore the procoagulant activity of factor VIII in the presence of the potent inhibitor antibody ESH8. J Biol Chem 2002; 277: 27232-9.
• 27 Lenting PJ, Donath M-JSH, van Mourik JA. et al. Identification of a binding site for blood coagulation factor IXa on the light chain of human factor VIII. J Biol Chem 1994; 269: 7150-5.
• 28 Healey JF, Barrow RT, Tamim HM. et al. Residues Glu2181 – Val2243 contain a major determinant of the inhibitory epitope in the C2 domain of human factor VIII. Blood 1998; 92: 3701-9.
• 29 Barrow RT, Healey JF, Jacquemin MG. et al. Antigenicity of putative phospholipid membrane-binding residues in factor VIII. Blood 2001; 97: 169-74.
• 30 Adams TE, Hockin MF, Mann KG. et al. The crystal structure of activated protein C-inactivated bovine factor Va: implications for cofactor function. Proc Natl Acad Sci U SA 2004; 101: 8918-23.
• 31 Ling M, Duncan EM, Rodgers SE. et al. Lowdetection rate of antibodies to non-functional epitopes on factor VIII in patients with hemophilia A and negative for inhibitors by Bethesda assay. J Thromb Haemost 2003; 1: 2548-53.
• 32 Klinge J, Auerswald G, Budde U. et al. Detection of all anti-factor VIII antibodies in haemophilia Apatients by the Bethesda assayand a more sensitive immunoprecipitation assay. Haemophilia 2001; 7: 26-32.
• 33 Dazzi F, Tison T, Vianello F. et al. High incidence of anti-FVIII antibodies against non-coagulant epitopes in haemophilia A patients: a possible role for the halflife of transfused FVIII. Br J Haematol 1996; 93: 688-93.