title: Platelet-derived ERp57 mediates platelet incorporation into a growing thrombus by regulation of the αIIbβ3 integrin.
abstract: The platelet protein disulfide isomerase called ERp57 mediates platelet aggregation, but its role in thrombus formation is unknown. To determine the specific role of platelet-derived ERp57 in hemostasis and thrombosis, we generated a megakaryocyte/platelet-specific knockout. Despite normal platelet counts and platelet glycoprotein expression, mice with ERp57-deficient platelets had prolonged tail-bleeding times and thrombus occlusion times with FeCl3-induced carotid artery injury. Using a mesenteric artery thrombosis model, we found decreased incorporation of ERp57-deficient platelets into a growing thrombus. Platelets lacking ERp57 have defective activation of the αIIbβ3 integrin and platelet aggregation. The defect in aggregation was corrected by the addition of exogenous ERp57, implicating surface ERp57 in platelet aggregation. Using mutants of ERp57, we demonstrate the second active site targets a platelet surface substrate to potentiate platelet aggregation. Binding of Alexa 488-labeled ERp57 to thrombin-activated and Mn(2+)-treated platelets lacking β3 was decreased substantially, suggesting a direct interaction of ERp57 with αIIbβ3. Surface expression of ERp57 protein and activity in human platelets increased with platelet activation, with protein expression occurring in a physiologically relevant time frame. In conclusion, platelet-derived ERp57 directly interacts with αIIbβ3 during activation of this receptor and is required for incorporation of platelets into a growing thrombus.
authors.affiliation: The Cyrus Tang Hematology Center, Jiangsu Institute of Hematology, First Affiliated Hospital, Soochow University, Suzhou, China;
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.forename: Yi
authors.initials: Y
authors.lastname: Wu
authors.forename: Junsong
authors.initials: J
authors.lastname: Zhou
authors.forename: Syed S
authors.initials: SS
authors.lastname: Ahmad
authors.forename: Bulent
authors.initials: B
authors.lastname: Mutus
authors.forename: Natalio
authors.initials: N
authors.lastname: Garbi
authors.forename: Günter
authors.initials: G
authors.lastname: Hämmerling
authors.forename: Junling
authors.initials: J
authors.lastname: Liu
authors.forename: David W
authors.initials: DW
authors.lastname: Essex
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000201:enzymology* / Q000502:physiology*; D020134:Catalytic Domain / Q000235:genetics; D006801:Humans; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000097:blood / Q000235:genetics; D015539:Platelet Activation / Q000502:physiology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000502:physiology*; D019704:Protein Disulfide-Isomerases / Q000097:blood* / Q000172:deficiency / Q000235:genetics; D011994:Recombinant Proteins / Q000097:blood / Q000235:genetics; D013927:Thrombosis / Q000097:blood* / Q000209:etiology
chemical_list: D050505:Mutant Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011994:Recombinant Proteins; C513195:Pdia3 protein, mouse; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human
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