title: The platelet glycoprotein Ib-von Willebrand factor interaction activates the collagen receptor alpha2beta1 to bind collagen: activation-dependent conformational change of the alpha2-I domain.
abstract: Integrin alpha2beta1 (glycoprotein [GP] Ia/IIa) is a major platelet receptor for collagen, containing its collagen-binding site within the alpha2 I domain. alpha2beta1 changes conformation upon platelet activation, increasing its affinity for collagen. We observed that 2 antibodies known to bind within the alpha2I domain, 12F1 and 6F1, bound preferentially to adenosine diphosphate (ADP)-activated platelets. Interestingly, when whole blood was perfused over a surface coated with either 12F1 or 6F1, only 6F1 supported the adhesion of unstimulated platelets. To test whether the interaction of GP Ib with von Willebrand factor (VWF) directly activates alpha2beta1, we used 12F1 as a probe of integrin activation. We perfused blood over a surface coated with a mixture of VWF-A1 domain (a GP Ib ligand) and 12F1 or VWF-A1 and mouse immunoglobulin G (IgG). Platelets rolled and did not attach stably on the A1/IgG surface, but they firmly bound and covered the A1/12F1 surface. We corroborated that 12F1 binds an active conformation of the I domain by showing that it binds with higher affinity to a gain-of-function mutant than to either wild-type I domain or a loss-of-function mutant. These results strongly suggest that the interaction of platelet GP Ib with VWF mediates the activation of alpha2beta1, increasing its affinity for collagen.
authors.affiliation: Thrombosis Research Section, Department of Medicine, Baylor College of Medicine, One Baylor Plaza, N1319, Houston, TX 77030, USA. miguelc@bcm.tmc.edu
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Cruz
authors.forename: Junmei
authors.initials: J
authors.lastname: Chen
authors.forename: Jody L
authors.initials: JL
authors.lastname: Whitelock
authors.forename: Liza D
authors.initials: LD
authors.lastname: Morales
authors.forename: José A
authors.initials: JA
authors.lastname: López
mesh_terms: D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D038982:Integrin alpha2beta1 / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D020125:Mutation, Missense; D010973:Platelet Adhesiveness; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D038981:Receptors, Collagen / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D038982:Integrin alpha2beta1; D019038:Platelet Glycoprotein GPIb-IX Complex; D038981:Receptors, Collagen; D014841:von Willebrand Factor; D003094:Collagen