title: Multimerin 1 binds factor V and activated factor V with high affinity and inhibits thrombin generation.
abstract: Multimerin 1 (MMRN1) is a polymeric, factor V (FV) binding protein that is stored in platelet and endothelial cell secretion granules but is undetectable in normal plasma. In human platelet alpha-granules, FV is stored complexed to MMRN1, predominantly by noncovalent binding interactions. The FV binding site for MMRN1 is located in the light chain, where it overlaps the C1 and C2 domain membrane binding sites essential for activated FV (FVa) procoagulant function. Surface plasmon resonance (SPR), circular dichroism (CD) and thrombin generation assays were used to study the binding of FV and FVa to MMRN1, and the functional consequences. FV and FVa bound MMRN1 with high affinities (K(D): 2 and 7 nM, respectively). FV dissociated more slowly from MMRN1 than FVa in SPR experiments, and CD analyses suggested greater conformational changes in mixtures of FV and MMRN1 than in mixtures of FVa and MMRN1. SPR analyses indicated that soluble phosphatidylserine (1,2-Dicaproylsn-glycero-3-phospho-L-serine) competitively inhibited both FV-MMRN1 and FVa-MMRN1 binding. Furthermore, exogenous MMRN1 delayed and reduced thrombin generation by plasma and platelets, and it reduced thrombin generation by preformed FVa. Exogenous MMRN1 also delayed FV activation, triggered by adding tissue factor to plasma, or by adding purified thrombin or factor Xa to purified FV. The high affinity binding of FV to MMRN1 may facilitate the costorage of the two proteins in platelet alpha-granules. As a consequence, MMRN1 release during platelet activation may limit platelet dependent thrombin generation in vivo.
authors.affiliation: Department of Pathology and Molecular Medicine, Health Sciences Center, McMaster University, 1200 Main St. West, Hamilton, Ontario, Canada, L8N 3Z5.
authors.forename: Samira B
authors.initials: SB
authors.lastname: Jeimy
authors.forename: Nola
authors.initials: N
authors.lastname: Fuller
authors.forename: Subia
authors.initials: S
authors.lastname: Tasneem
authors.forename: Kenneth
authors.initials: K
authors.lastname: Segers
authors.forename: Alan R
authors.initials: AR
authors.lastname: Stafford
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Weitz
authors.forename: Rodney M
authors.initials: RM
authors.lastname: Camire
authors.forename: Gerry A F
authors.initials: GA
authors.lastname: Nicolaes
authors.forename: Catherine P M
authors.initials: CP
authors.lastname: Hayward
mesh_terms: D001667:Binding, Competitive; D001792:Blood Platelets / Q000378:metabolism*; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D005165:Factor V / Q000737:chemistry / Q000378:metabolism*; D015943:Factor Va / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010718:Phosphatidylserines / Q000097:blood; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013917:Thrombin / Q000378:metabolism*
chemical_list: C402786:1,2-dicaproylphosphatidylserine; D001798:Blood Proteins; D010718:Phosphatidylserines; D011994:Recombinant Proteins; C069282:multimerin; D015943:Factor Va; D005165:Factor V; D013917:Thrombin; D015951:Factor Xa