Department / Division:
Medicine / Hematology

Address:
DUMC 3656
Durham, NC 27710

Appointment Telephone:
919-668-6688

Office Telephone:
919-684-5350

Fax Telephone:
919-681-6160

• MD, PhD, Washington University in St. Louis School of Medicine (Missouri), 1982

• Medicine, Barnes Hospital (Missouri), 1982-1984

• Hematology/Oncology, Barnes Hospital (Missouri), 1984-1985

Clinical Interests:
Coagulation factor inhibitors and deficiencies, recurrent thrombosis, hypercoagulable states, protein C resistance, lupus anticoagulants, anticoagulation management, pre-operative hemostasis evaluation, von Willebrand disease, hemophilia, diagnosis and treatment of complex coagulation problems

Research Interests:
In the United States, cardiovascular disease results in one death every 30 seconds.  Clinical disorders such as myocardial infarction, deep vein thrombosis and pulmonary embolism, and stroke are usually precipitated by thrombotic events.  Although basic research in thrombosis has lead to significant advances in the diagnosis and treatment of thrombotic disorders current approaches remain sub optimal.  

Generation of thrombin by the prothrombinase complex plays a particularly important role in the pathogenesis venous thrombosis.  The prothrombinase complex consists of the enzyme factor Xa, the cofactor factor Va and a phospholipid membrane surface.  The interaction of factor Xa with factor Va requires cofactor activation for expression of factor Xa binding sites.  The interaction of factor Va with platelet membranes requires expression of phosphatidylserine (PS) on the surface of activated platelets or endothelial cells.  The binding sites for factor Xa and phospholipid membranes are discontinuous and located in several different domains.  The complexity of these binding sites may allow for the fine regulation of the prothrombinase complex. The molecular bases for these interactions remain poorly understood.    

The long-term goal of this project is to use integrated molecular, structural and biophysical approaches to understand the interaction of factor Va with biological membranes.    We have expressed the factor V C2 domain using insect cells and determined the structures of two crystal forms.  We have also used factor Va mutants expressed in mammalian cells to define the roles of solvent exposed hydrophobic amino acid residues in membrane binding and prothrombinase assembly.   Two tryptophans located in a mobile solvent exposed loop play a critical role in high affinity binding of factor V to phospholipid membranes containing low concentrations of PS.  A second membrane-binding site, which also requires a pair of solvent exposed hydrophobic amino acid residues, has recently been localized in the factor V C1 domain.  We are also collaborating with Dr. Barry Lentz at the University of North Carolina to study the interaction of factor Va mutants with soluble phosphatidyl serine (C6PS).  Recent studies by Dr. Lentz’s group have demonstrated that a fully functional prothrombinase complex can be assembled in solution in the presence of C6PS suggesting that PS is an allosteric regulator of factor Va cofactor activity.

The goals of our present research are to further define the binding sites in the factor Va light chain for phospholipid membranes, C6PS and cellular membranes. Binding sites will be localized using recombinant factor Va mutants, recombinant light chain domains, domain specific and monoclonal antibodies.    Experiments will be designed using available crystal structures or molecular models for individual domains.  Binding interactions will be characterized using surface plasmon resonance and fluorescence binding assays.   This information will provide important new insights into regulation of the prothrombinase complex including the molecular basis for specific high affinity binding of factor Va to PS membranes and regulation of cofactor activity by PS.  This knowledge could lead to novel targets for anti-thrombotic therapy.

Representative Publications:
Peng W, Quinn-Allen MA, Kane WH. Mutation of hydrophobic residues in the factor Va C1 and C2 domains blocks membrane-dependent prothrombin activation. J Thromb Haemost. 2005 Feb;3(2):351-4. (2005) Abstract

Majumder R, Quinn-Allen MA, Kane WH, Lentz BR. The Phosphatidylserine Binding Site of the Factor V(a) C2 Domain Accounts for Membrane Binding but Does Not Contribute to the Assembly or Activity of a Human Factor X(a)-Factor V(a) Complex. Biochemistry. 2005 Jan 18;44(2):711-8. (2005) Abstract

Saleh M, Peng W, Quinn-Allen MA, Macedo-Ribeiro S, Fuentes-Prior P, Bode W, Kane WH. The factor V C1 domain is involved in membrane binding: identification of functionally important amino acid residues within the C1 domain of factor V using alanine scanning mutagenesis.  Thromb Haemost.  2004 Jan;91(1):16-27. (2004) Abstract

Peng W, Quinn-Allen MA, Kim SW, Alexander KA, Kane WH. Trp2063 and trp2064 in the factor Va C2 domain are required for high-affinity binding to phospholipid membranes but not for assembly of the prothrombinase complex.  Biochemistry.  2004 Apr 13;43(14):4385-93. (2004) Abstract

Jeimy SB, Woram RA, Fuller N, Quinn-Allen MA, Nicolaes GA, Dahlbäck B, Kane WH, Hayward CP. Identification of the MMRN1 Binding Region within the C2 Domain of Human Factor V. J Biol Chem. 2004 Dec 3;279(49):51466-71. (2004) Abstract

Hayward CP, Fuller N, Zheng S, Adam F, Jeimy SB, Horsewood I, Ann Quinn-Allen M, Kane WH. Human platelets contain forms of factor V in disulfide-linkage with multimerin.  Thromb Haemost.  2004 Dec;92(6):1349-57. (2004) Abstract

Srivastava A, Quinn-Allen MA, Kim SW, Kane WH, Lentz BR. Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor Va.  Biochemistry.  2001 Jul 27;40(28):8246-55. (2001) Abstract

Izumi T, Kim SW, Greist A, Macedo-Ribeiro S, Fuentes-Prior P, Bode W, Kane WH, Ortel TL. Fine mapping of inhibitory anti-factor V antibodies using factor V C2 domain mutants. Identification of two antigenic epitopes involved in phospholipid binding.  Thromb Haemost.  2001 Jun;85(6):1048-54. (2001) Abstract

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 Feb 29;39(8):1951-8. (2000) Abstract

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 Nov 25;402(6760):434-9. (1999) Abstract

Kim SW, Ortel TL, Quinn-Allen MA, Yoo L, Worfolk L, Zhai X, Lentz BR, Kane WH. Partial glycosylation at asparagine-2181 of the second C-type domain of human factor V modulates assembly of the prothrombinase complex.  Biochemistry.  1999 Aug 31;38(35):11448-54. (1999) Abstract

Ortel TL, Moore KD, Quinn-Allen MA, Okamura T, Sinclair AJ, Lazarchick J, Govindan R, Carmagnol F, Kane WH. Inhibitory anti-factor V antibodies bind to the factor V C2 domain and are associated with hemorrhagic manifestations.  Blood.  1998 Jun 1;91(11):4188-96. (1998) Abstract

Hayward CP, Cramer EM, Kane WH, Zheng S, Bouchard M, Massé JM, Rivard GE. Studies of a second family with the Quebec platelet disorder: evidence that the degradation of the alpha-granule membrane and its soluble contents are not secondary to a defect in targeting proteins to alpha-granules. Blood. 1997 Feb 15;89(4):1243-53. (1997) Abstract

Keller FG, Ortel TL, Quinn-Allen MA, Kane WH. Thrombin-catalyzed activation of recombinant human factor V.  Biochemistry.  1995 Mar 28;34(12):4118-24. (1995) Abstract

Ortel TL, Quinn-Allen MA, Charles LA, Devore-Carter D, Kane WH. Characterization of an acquired inhibitor to coagulation factor V. Antibody binding to the second C-type domain of factor V inhibits the binding of factor V to phosphatidylserine and neutralizes procoagulant activity.  J Clin Invest.  1992 Dec;90(6):2340-7. (1992) Abstract

Cripe LD, Moore KD, Kane WH. Structure of the gene for human coagulation factor V.  Biochemistry.  1992 Apr 21;31(15):3777-85. (1992) Abstract

Kane WH, Davie EW. Blood coagulation factors V and VIII: structural and functional similarities and their relationship to hemorrhagic and thrombotic disorders.  Blood.  1988 Mar;71(3):539-55. (1988) Abstract

Kane WH, Ichinose A, Hagen FS, Davie EW. Cloning of cDNAs coding for the heavy chain region and connecting region of human factor V, a blood coagulation factor with four types of internal repeats.  Biochemistry.  1987 Oct 6;26(20):6508-14. (1987) Abstract

Kane WH, Davie EW. Cloning of a cDNA coding for human factor V, a blood coagulation factor homologous to factor VIII and ceruloplasmin.  Proc Natl Acad Sci U S A.  1986 Sep;83(18):6800-4. (1986) Abstract

Kane WH, Majerus PW. The interaction of human coagulation factor Va with platelets.  J Biol Chem.  1982 Apr 10;257(7):3963-9. (1982) Abstract

Kane WH, Majerus PW. Purification and characterization of human coagulation factor V.  J Biol Chem.  1981 Jan 25;256(2):1002-7. (1981) Abstract