A point mutation in the integrin beta 3 cytoplasmic domain (S752--> P) impairs bidirectional signaling through alpha IIb beta 3 (platelet glycoprotein IIb-IIIa)

YP Chen, TE O'Toole, J Ylanne, JP Rosa, MH Ginsberg - 1994 - ashpublications.org
YP Chen, TE O'Toole, J Ylanne, JP Rosa, MH Ginsberg
1994ashpublications.org
Agonist-induced inside-out signaling results in an increased affinity of integrin alpha IIb beta
3 (platelet glycoprotein IIb-IIIa) for soluble ligands (fibrinogen [Fg] and PAC1). Ligand
binding to integrins initiates outside-in signaling that leads to cellular responses such as cell
spreading and focal adhesion formation. A point mutation in the beta 3 cytoplasmic domain
(S752--> P) is associated with blocked inside-out alpha IIb beta 3 signaling in a variant
Glanzmann's thrombasthenia. This mutation was introduced into beta 3 and cotransfected …
Abstract
Agonist-induced inside-out signaling results in an increased affinity of integrin alpha IIb beta 3 (platelet glycoprotein IIb-IIIa) for soluble ligands (fibrinogen [Fg] and PAC1). Ligand binding to integrins initiates outside-in signaling that leads to cellular responses such as cell spreading and focal adhesion formation. A point mutation in the beta 3 cytoplasmic domain (S752-->P) is associated with blocked inside- out alpha IIb beta 3 signaling in a variant Glanzmann's thrombasthenia. This mutation was introduced into beta 3 and cotransfected into Chinese hamster ovary cells with a chimeric alpha subunit consisting of the alpha IIb extracellular and transmembrane domains and the alpha 6B cytoplasmic domain. The substitution of the alpha IIb cytoplasmic domain with that of alpha 6 led to activation of alpha IIb beta 3 to bind PAC1, mimicking inside-out signaling. This effect was reversed by the S752-->P mutation, indicating a disruption of inside-out signaling by the mutation. In addition, transfectants expressing this beta 3 variant showed reduced alpha IIb beta 3-mediated cell spreading on immobilized Fg, focal adhesion, and fibrin clot retraction, suggesting an impairment in outside-in alpha IIb beta 3 signaling. Therefore, a single point mutation in the beta 3 cytoplasmic domain impaired bidirectional signaling through alpha IIb beta 3.
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