title: Coated platelets function in platelet-dependent fibrin formation via integrin αIIbβ3 and transglutaminase factor XIII.
abstract: Coated platelets, formed by collagen and thrombin activation, have been characterized in different ways: i) by the formation of a protein coat of α-granular proteins; ii) by exposure of procoagulant phosphatidylserine; or iii) by high fibrinogen binding. Yet, their functional role has remained unclear. Here we used a novel transglutaminase probe, Rhod-A14, to identify a subpopulation of platelets with a cross-linked protein coat, and compared this with other platelet subpopulations using a panel of functional assays. Platelet stimulation with convulxin/thrombin resulted in initial integrin α(IIb)β3 activation, the appearance of a platelet population with high fibrinogen binding, (independently of active integrins, but dependent on the presence of thrombin) followed by phosphatidylserine exposure and binding of coagulation factors Va and Xa. A subpopulation of phosphatidylserine-exposing platelets bound Rhod-A14 both in suspension and in thrombi generated on a collagen surface. In suspension, high fibrinogen and Rhod-A14 binding were antagonized by combined inhibition of transglutaminase activity and integrin α(IIb)β3 Markedly, in thrombi from mice deficient in transglutaminase factor XIII, platelet-driven fibrin formation and Rhod-A14 binding were abolished by blockage of integrin α(IIb)β3. Vice versa, star-like fibrin formation from platelets of a patient with deficiency in α(IIb)β3(Glanzmann thrombasthenia) was abolished upon blockage of transglutaminase activity. We conclude that coated platelets, with initial α(IIb)β3 activation and high fibrinogen binding, form a subpopulation of phosphatidylserine-exposing platelets, and function in platelet-dependent star-like fibrin fiber formation via transglutaminase factor XIII and integrin α(IIb)β3.
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Nadine J A
authors.initials: NJ
authors.lastname: Mattheij
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Frauke
authors.initials: F
authors.lastname: Swieringa
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Tom G
authors.initials: TG
authors.lastname: Mastenbroek
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, Portland, OR, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Berny-Lang
authors.affiliation: CSL Behring GmbH, Marburg, Germany.
authors.forename: Frauke
authors.initials: F
authors.lastname: May
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Constance C F M J
authors.initials: CC
authors.lastname: Baaten
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Paola E J
authors.initials: PE
authors.lastname: van der Meijden
authors.affiliation: Central Diagnostic Laboratory, Maastricht University Medical Center, The Netherlands.
authors.forename: Yvonne M C
authors.initials: YM
authors.lastname: Henskens
authors.affiliation: Department of Internal Medicine, Maastricht University Medical Center, The Netherlands.
authors.forename: Erik A M
authors.initials: EA
authors.lastname: Beckers
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Dennis P L
authors.initials: DP
authors.lastname: Suylen
authors.affiliation: CSL Behring GmbH, Marburg, Germany.
authors.forename: Marc W
authors.initials: MW
authors.lastname: Nolte
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Tilman M
authors.initials: TM
authors.lastname: Hackeng
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, Portland, OR, USA.
authors.forename: Owen J T
authors.initials: OJ
authors.lastname: McCarty
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
authors.forename: Johan W M
authors.initials: JW
authors.lastname: Heemskerk
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands judith.cosemans@maastrichtuniversity.nl.
authors.forename: Judith M E M
authors.initials: JM
authors.lastname: Cosemans
mesh_terms: D000818:Animals; D001777:Blood Coagulation; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D003435:Crotalid Venoms / Q000494:pharmacology; D015943:Factor Va / Q000737:chemistry / Q000378:metabolism; D005176:Factor XIII / Q000737:chemistry / Q000378:metabolism*; D015951:Factor Xa / Q000737:chemistry / Q000378:metabolism; D005337:Fibrin / Q000737:chemistry / Q000378:metabolism*; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type; D051379:Mice; D018345:Mice, Knockout; D015335:Molecular Probes / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism; D015539:Platelet Activation / Q000187:drug effects; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry / Q000378:metabolism*; D061251:Primary Cell Culture; D011485:Protein Binding; D013915:Thrombasthenia / Q000097:blood* / Q000473:pathology; D013917:Thrombin / Q000494:pharmacology
chemical_list: D003435:Crotalid Venoms; D037181:Lectins, C-Type; D015335:Molecular Probes; D010718:Phosphatidylserines; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; C028890:convulxin; D015943:Factor Va; D005337:Fibrin; D005340:Fibrinogen; D005176:Factor XIII; D013917:Thrombin; D015951:Factor Xa