title: Outside-in signaling from integrin alpha IIb beta 3 into platelets in the absence of agonist-induced signaling.
abstract: Platelet agonists generate intracellular signals which lead to activation of the platelet membrane glycoprotein IIb-IIIa (integrin alpha IIb beta 3). The resulting occupancy of alpha IIb beta 3 by ligands also generates signals into the cell (outside-in signaling). We reported previously that unlike platelet agonists, the F(ab')2 fragments of an anti-alpha IIb beta 3 monoclonal antibody, PMA4, induced fibrinogen binding to alpha IIb beta 3 without causing intracellular activation. In this study, in order to determine whether outside-in signaling occurs in the absence of agonist-induced intracellular signals, we used PMA4 F(ab')2 as an inducer of fibrinogen binding to alpha IIb beta 3. PMA4 F(ab')2-induced fibrinogen binding and subsequent platelet aggregation triggered tyrosine phosphorylation of several proteins including pp72syk but not pp125FAK. No Ca2+ influx or mobilization, thromboxane B2 synthesis, phosphorylation of pleckstrin or the myosin light chain, cytoplasmic alkalinization, or platelet shape changes, were detected. These findings suggest that, in the absence of agonist-induced signaling, alpha IIb beta 3 occupied by soluble fibrinogen generates only a limited outside-in signal.
authors.affiliation: Blood Transfusion Division, Ehime University School of Medicine, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Hato
authors.forename: A
authors.initials: A
authors.lastname: Oda
authors.forename: Y
authors.initials: Y
authors.lastname: Ozaki
authors.forename: Y
authors.initials: Y
authors.lastname: Minamoto
authors.forename: S
authors.initials: S
authors.lastname: Nakatani
authors.forename: A
authors.initials: A
authors.lastname: Watanabe
authors.forename: S
authors.initials: S
authors.lastname: Kume
authors.forename: Y
authors.initials: Y
authors.lastname: Ikeda
authors.forename: S
authors.initials: S
authors.lastname: Fujita
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000097:blood; D015815:Cell Adhesion Molecules / Q000097:blood; D004792:Enzyme Precursors / Q000097:blood; D005340:Fibrinogen / Q000378:metabolism; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D047908:Intracellular Signaling Peptides and Proteins; D010766:Phosphorylation; D015539:Platelet Activation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000097:blood; D015398:Signal Transduction; D000072377:Syk Kinase; D013929:Thromboxane B2 / Q000097:blood; D014443:Tyrosine / Q000097:blood
chemical_list: D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D004792:Enzyme Precursors; D007140:Immunoglobulin Fab Fragments; D047908:Intracellular Signaling Peptides and Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D014443:Tyrosine; D013929:Thromboxane B2; D005340:Fibrinogen; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human; C000606896:SYK protein, human; D000072377:Syk Kinase; D002118:Calcium