title: Glutamate Receptor Interacting Protein 1 Mediates Platelet Adhesion and Thrombus Formation.
abstract: Thrombosis-associated pathologies, such as myocardial infarction and stroke, are major causes of morbidity and mortality worldwide. Because platelets are necessary for hemostasis and thrombosis, platelet directed therapies must balance inhibiting platelet function with bleeding risk. Glutamate receptor interacting protein 1 (GRIP1) is a large scaffolding protein that localizes and organizes interacting proteins in other cells, such as neurons. We have investigated the role of GRIP1 in platelet function to determine its role as a molecular scaffold in thrombus formation. Platelet-specific GRIP1-/- mice were used to determine the role of GRIP1 in platelets. GRIP1-/- mice had normal platelet counts, but a prolonged bleeding time and delayed thrombus formation in a FeCl3-induced vessel injury model. In vitro stimulation of WT and GRIP1-/- platelets with multiple agonists showed no difference in platelet activation. However, in vivo platelet rolling velocity after endothelial stimulation was significantly greater in GRIP1-/- platelets compared to WT platelets, indicating a potential platelet adhesion defect. Mass spectrometry analysis of GRIP1 platelet immunoprecipitation revealed enrichment of GRIP1 binding to GPIb-IX complex proteins. Western blots confirmed the mass spectrometry findings that GRIP1 interacts with GPIbα, GPIbβ, and 14-3-3. Additionally, in resting GRIP1-/- platelets, GPIbα and 14-3-3 have increased interaction compared to WT platelets. GRIP1 interactions with the GPIb-IX binding complex are necessary for normal platelet adhesion to a stimulated endothelium.
authors.affiliation: Aab Cardiovascular Research Institute, University of Rochester Medical Center, Rochester, NY, United States of America.|Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, NY, United States of America.
authors.forename: Kristina L
authors.initials: KL
authors.lastname: Modjeski
authors.affiliation: Aab Cardiovascular Research Institute, University of Rochester Medical Center, Rochester, NY, United States of America.
authors.forename: Sara K
authors.initials: SK
authors.lastname: Ture
authors.affiliation: Aab Cardiovascular Research Institute, University of Rochester Medical Center, Rochester, NY, United States of America.
authors.forename: David J
authors.initials: DJ
authors.lastname: Field
authors.affiliation: Aab Cardiovascular Research Institute, University of Rochester Medical Center, Rochester, NY, United States of America.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Cameron
authors.affiliation: Aab Cardiovascular Research Institute, University of Rochester Medical Center, Rochester, NY, United States of America.
authors.forename: Craig N
authors.initials: CN
authors.lastname: Morrell
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D010973:Platelet Adhesiveness / Q000502:physiology*
chemical_list: D002352:Carrier Proteins; C491310:GRIP1 protein, human; D009419:Nerve Tissue Proteins