Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1995-5-30
pubmed:abstractText
The affinity of integrin alpha IIb beta 3 for adhesive ligands is tightly regulated by the platelet such that fibrinogen binding is observed only after platelet activation. Ligand binding is necessary for platelet aggregation, which contributes to vascular occlusion in pathological states. Therefore, we have developed an ELISA assay to screen for compounds that inhibit alpha IIb beta 3 activation. Washed platelets were incubated in microtitre wells with potential inhibitory compounds and stimulated with an agonist to activate alpha IIb beta 3. After the addition of biotin-PAC1, a fibrinogen-mimetic monoclonal antibody, the activation state of alpha IIb beta 3 was measured by sedimenting the platelets and quantitating the residual biotin-PAC1 in the cell-free supernatant in a streptavidin-based ELISA. This assay detected (1) specific PAC1 binding to activated platelets in response to a variety of agonists, and (2) dose-dependent inhibition of PAC1 binding by function-blocking anti-alpha IIb beta 3 monoclonal antibodies, by the tetrapeptide, RGDS, and by an alpha IIb beta 3-selective RGD peptidomimetic. Furthermore, the assay detected inhibition of PAC1 binding by intracellular inhibitors of platelet activation, including bisindolylmaleimide, a selective protein kinase C antagonist, and wortmannin, an inhibitor of phosphatidylinositol 3-kinase. These studies demonstrate that this integrin activation ELISA can detect pharmacological blockade of platelet alpha IIb beta 3 by extracellular and intracellular inhibitors. Its use may facilitate the search for clinically useful anti-platelet drugs.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0022-1759
pubmed:author
pubmed:issnType
Print
pubmed:day
12
pubmed:volume
181
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
55-64
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1995
pubmed:articleTitle
An enzyme-linked immunosorbent assay to identify inhibitors of activation of platelet integrin alpha IIb beta 3.
pubmed:affiliation
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't