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Prepublished online as a Blood First Edition Paper on May 24, 2002; DOI 10.1182/blood-2002-02-0418. This Article Full Text (PDF) All Versions of this Article: 2002-02-0418v1 100/6/2094    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Sun, Q.-H. Articles by Newman, P. J Search for Related Content PubMed PubMed Citation Articles by Sun, Q.-H. Articles by Newman, P. J Social Bookmarking                   What's this? Submitted February 8, 2002 Accepted May 6, 2002 Disruption of the long-range GPIIIa Cys5-Cys435 disulfide bond results in the production of constitutively active GPIIb-IIIa (IIbß3) integrin complexes Qi-Hong Sun*, Chao-Yan Liu, Ronggang Wang, Cathy Paddock, and Peter J Newman Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, WI, USA; Cardiovascular Center, The Medical College of Wisconsin, Milwaukee, WI, USA Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, WI, USA Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, WI, USA; Departments of Cellular Biology and Pharmacology, The Medical College of Wisconsin, Milwaukee, WI, USA * Corresponding author; email: qsun{at}bcsew.edu. The major platelet integrin IIbß3, also known as the platelet glycoprotein (GP) IIb-IIIa complex, mediates platelet aggregation by serving as the receptor for fibrinogen and von Willebrand factor. In addition to its physiologic role, GPIIb-IIIa also bears a number of clinically important alloantigenic determinants. Previous studies have shown that disruption of the long-range Cys5-Cys435 disulfide bond of the ß3 subunit results in the production of GPIIIa isoforms that bind some, but not all, anti-PlA1 alloantibodies, suggesting that mutations in this so-called long-range disulfide bond can alter the conformation of GPIIIa. The purpose of this study was to examine the effect of either the Cys5Ala or Cys435Ala substitution of GPIIIa on the adhesive properties of the GPIIb-IIIa complex. We found that both Ala5GPIIIa and Ala435GPIIIa were capable of associating with GPIIb and were expressed normally on the cell surface when co-transfected into Chinese Hamster Ovary (CHO) cells. CHO cells expressing GPIIb-Ala5GPIIIa or GPIIb-Ala435IIIa bound well-characterized, conformationally-sensitive LIBS (ligand-induced binding site) antibodies, and were capable of constitutively binding the fibrinogen-mimetic monoclonal antibodies PL-55 and PAC-1, as well as soluble fibrinogen. Both GPIIb-Ala5IIIa and GPIIb-Ala435IIIa-transfected CHO cells also bound more avidly to immobilized fibrinogen, and were capable of mediating the tyrosine phosphorylation of pp125FAK upon cell adhesion. These data are consistent with the notion that these regions of GPIIIa participate in the conformational change associated with receptor activation. Additionally, these studies may provide a molecular explanation for the previously reported ability of mild reducing agents to activate the GPIIb-IIIa complex and promote platelet aggregation. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: M. Kannan and R. 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