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Thromb Haemost 2004; 92(06): 1349-1357
DOI: 10.1160/TH03-02-0123
Platelets and Blood Cells
Schattauer GmbH

Human platelets contain forms of factor V in disulfide-linkage with multimerin

Catherine P. M. Hayward
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
Nola Fuller
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
Shilun Zheng
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
Frédéric Adam
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
Samira B. Jeimy
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
Ian Horsewood
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
Mary Ann Quinn-Allen
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
,
William H. Kane
1   Departments of Pathology and Molecular Medicine, and Medicine, McMaster University, and Hamilton Health Sciences and the Division of Hematology-Oncology and Departments of Medicine and Pathology, Duke University Medical Centre, Durham, North Carolina, USA
› Author AffiliationsFinancial support: Supported by grant 42450 from the Canadian Institutes of Health Research (C.P.M. Hayward) and grants HL43106 and HL54939 from the National Institutes of Health (W.H. Kane). C.P.M. Hayward is the recipient of a Career Investigator Award from the Heart and Stroke Foundation of Ontario and a Canada Research Chair in Molecular Hemostasis. Samira B. Jeimy is the recipient of a Heart and Stroke Foundation of Ontario Research Master’s Studentship Award.
Further Information

Publication History

Received 27 February 2003

Accepted after revision 17 September 2004

Publication Date:
02 December 2017 (online)

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Summary

Factor V is an essential cofactor for blood coagulation that circulates in platelets and plasma. Unlike plasma factor V, platelet factorV is stored complexed with the polymeric α-granule protein multimerin. In analyses of human platelet factor V on nonreduced denaturing multimer gels, we identified that approximately 25% was variable in size and migrated larger than single chain factor V, the largest form in plasma. Upon reduction, the unusually large, variably-sized forms of platelet factor V liberated components that comigrated with other forms of platelet factor V, indicating that they contained factor V in interchain disulfide-linkages. With thrombin cleavage, factor Va heavy and light chain domains, but not B-domains, were liberated from the components linked by interchain disulfide bonds, indicating that the single cysteine in the B-domain at position 1085 was the site of disulfide linkage. Since unusually large factor V had a variable size and included forms larger than factor V dimers, the data suggested disulfide-linkage with another platelet protein, possibly multimerin. Immunoprecipitation experiments confirmed that unusually large factor V was associated with multimerin and it remained associated in 0.5 M salt. Moreover, platelets contained a subpopulation of multimerin polymers that resisted dissociation from factor V by denaturing detergent and comigrated with unusually large platelet factor V, before and after thrombin cleavage.The disulfide-linked complexes of multimerin and factor V in platelets, which are cleaved by thrombin to liberate factor Va, could be important for modulating the function of platelet factor V and its delivery onto activated platelets. Factor Va generation and function from unusually large platelet factor V is only speculative at this time.

Keywords

Factor V - multimerin - coagulation - platelets - alpha-granule proteins
 

  • References

  • 1 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.
    PubMedSearch in Google Scholar
  • 2 Mann KG, Kalafatis M. Factor V: a combination of Dr Jekyll and Mr. Hyde. Blood 2003; 101: 20-30.
    CrossrefPubMedSearch in Google Scholar
  • 3 Cui J, O’Shea S, Purkayastha A. et al. Fatal hemorrhage and incomplete block to embryogenesis in mice lacking coagulation factor V. Nature 1996; 384: 66-8.
    CrossrefPubMedSearch in Google Scholar
  • 4 Miletich JP, Majerus DW, Majerus PW. Patients with congenital factor V deficiency have decreased factor Xa binding sites on their platelets. J Clin Invest 1978; 62: 824-31.
    CrossrefPubMedSearch in Google Scholar
  • 5 Guasch JF, Cannegieter S, Reitsma PH. et al. Severe coagulation factor V deficiency caused by a 4 bp deletion in the factor V gene. Br J Haematol 1998; 101: 32-9.
    CrossrefPubMedSearch in Google Scholar
  • 6 Chediak J, Ashenhurst JB, Garlick I. et al. Successful management of bleeding in a patient with factor V inhibitor by platelet transfusions. Blood 1980; 56: 835-41.
    PubMedSearch in Google Scholar
  • 7 Borchgrevink CF, Owren PA. The hemostatic effect of normal platelets in hemophilia and factor V deficiency: the importance of clotting factors adsorbed on platelets for normal hemostasis. Acta Med Scand 1961; 170: 375-83.
    PubMedSearch in Google Scholar
  • 8 Nesheim ME, Nichols WL, Cole TL. et al. Isolation and study of an acquired inhibitor of human coagulation factor V. J Clin Invest 1986; 77: 405-15.
    CrossrefPubMedSearch in Google Scholar
  • 9 Cripe LD, Moore KD, Kane WH. Structure of the gene for human coagulation factor V. Biochemistry 1992; 31: 3777-85.
    CrossrefPubMedSearch in Google Scholar
  • 10 Viskup RW, Tracy PB, Mann KG. Isolation of human platelet factor V. Blood 1987; 69: 1188-95.
    PubMedSearch in Google Scholar
  • 11 Gould WR, Silveira JR, Tracy PB. Unique in vivo modifications of coagulation factor V produce a physically and functionally distinct platelet-derived cofactor: characterization of purified platelet-derived factor V/Va. J Biol Chem 2004; 279: 2383-93.
    CrossrefPubMedSearch in Google Scholar
  • 12 Hayward CPM, Furmaniak-Kazmierczak E, Cieutat A-M. et al. Factor V is complexed with multimerin in resting platelet lysates and colocalizes with multimerin in platelet α-granules. J Biol Chem 1995; 270: 19217-24.
    CrossrefPubMedSearch in Google Scholar
  • 13 Hayward CPM. Multimerin: a bench to bedside chronology of a unique platelet and endothelial cell protein - from discovery to function to abnormalities in disease. Clin Invest Med 1997; 20: 176-87.
    PubMedSearch in Google Scholar
  • 14 Hayward CPM, Weiss HJ, Lages B. et al. The storage defects in grey platelet syndrome and αδ-storage pool deficiency affect α-granule factor V and multimerin storage without altering their proteolytic processing. Br J Haematol 2001; 113: 871-7.
    CrossrefPubMedSearch in Google Scholar
  • 15 Hayward CPM, Rivard GE, Kane WH. et al. An autosomal dominant, qualitative platelet disorder associated with multimerin deficiency, abnormalities in platelet factor V, thrombospondin, von Willebrand factor, and fibrinogen and an epinephrine aggregation defect. Blood 1996; 87: 4967-78.
    PubMedSearch in Google Scholar
  • 16 Hayward CPM, Smith JW, Horsewood P. et al. P-155, a multimeric platelet protein that is expressed on activated platelets. J Biol Chem 1991; 266: 7114-20.
    PubMedSearch in Google Scholar
  • 17 Kahr WHA, Zheng S, Sheth PM. et al. Platelets from patients with the Quebec platelet disorder contain and secrete abnormal amounts of urokinase type plasminogen activator. Blood 2001; 98: 257-65.
    CrossrefPubMedSearch in Google Scholar
  • 18 Riddles PW, Blakeley RL, Zerner B. Reassessment of Ellman’s reagent. Methods Enzymol 1983; 91: 49-60.
    PubMedSearch in Google Scholar
  • 19 Srivastava A, Quinn-Allen MA, Kim SW. et al. Soluble phosphatidylserine binds to a single identified site in the C2 domain of human factor Va. Biochemistry 2001; 40: 8246-55.
    CrossrefPubMedSearch in Google Scholar
  • 20 Harlow E, Lane D. Antibodies: A Laboratory Manual. Cold Spring Harbor, Cold Spring Harbor Laboratory. 1988
    PubMedSearch in Google Scholar
  • 21 Horsewood P, Hayward CP, Warkentin TE. et al. Investigation of the mechanisms of monoclonal antibody-induced platelet activation. Blood 1991; 78: 1019-26.
    PubMedSearch in Google Scholar
  • 22 Hayward CPM, Song Z, Zheng S. et al. Multimerin processing by cells with and without pathways for regulated protein secretion. Blood 1999; 94: 1337-47.
    PubMedSearch in Google Scholar
  • 23 Hayward CPM, Warkentin TE, Horsewood P. et al. Multimerin: a series of large, disulfidelinked multimeric proteins within platelets. Blood 1991; 77: 2556-60.
    PubMedSearch in Google Scholar
  • 24 Kane WH, Ichinose A, Hagen FS. et al. 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; 26: 6508-14.
    CrossrefPubMedSearch in Google Scholar
  • 25 Jenny RJ, Pittman DD, Toole JJ. et al. Complete cDNA and derived amino acid sequence of human factor V. Proc Natl Acad Sci 1987; 84: 4846-50.
    CrossrefPubMedSearch in Google Scholar
  • 26 Mannucci PM. Platelet von Willebrand factor in inherited and acquired bleeding disorders. Proc Natl Acad Sci 1995; 92: 2428-32.
    CrossrefPubMedSearch in Google Scholar
  • 27 Monkovic DD, Tracy PB. Functional characterization of human platelet-released factor V and and its activation by factor Xa and thrombin. J Biol Chem 1990; 265: 17132-40.
    PubMedSearch in Google Scholar
  • 28 Essex DW, Chen K, Swiatkowska M. Localization of protein disulfide isomerase to the external surface platelet membrane. Blood 1995; 86: 2168-73.
    PubMedSearch in Google Scholar
  • 29 Thorelli E, Kaufman RJ, Dahlback B. The Cterminal region of the factor V BD is crucial for the anticoagulant activity of factor V. J Biol Chem 1998; 273: 16140-5.
    CrossrefPubMedSearch in Google Scholar
  • 30 Guinto ER, Esmon CT, Mann KG. et al. The complete cDNA sequence of bovine coagulation factor V. J Biol Chem 1992; 267: 2971-8.
    PubMedSearch in Google Scholar
  • 31 Yang TL, Cui J, Rehumtulla A. et al. The structure and function of murine factor V and its inactivation by protein C. Blood 1998; 91: 4593-9.
    PubMedSearch in Google Scholar
  • 32 Wyshock EG, Stewart GJ, Colman RW. Thrombin-stimulated human platelets express molecular forms of α-granular factor V (FV), which differ from FVa. Thromb Haemost 1994; 72: 947-56.
    Thieme ConnectPubMedSearch in Google Scholar