title: Redox control of platelet aggregation.
abstract: Sulfhydryl and disulfide metabolism in platelet function has recently reemerged as a focus of platelet research. In this study we tested the effect of redox buffer on platelet aggregation and the effect of reduced glutathione (GSH) and platelet activation on sulfhydryl exposure in the platelet fibrinogen receptor, alpha IIb beta 3. In the presence of subthreshold concentrations of agonist, physiologic concentrations of GSH (10 microM) stimulated platelet aggregation and secretion. These effects were found with more than one platelet agonist and with different low molecular weight thiols, including homocysteine. The effect of low molecular weight thiols was reproduced with the peptide LSARLAF which directly activates platelets through alpha IIb beta 3, suggesting that the mechanism is at the level of this integrin. After determining optimal sulfhydryl labeling conditions for alpha IIb beta 3 (5 mM EDTA, 37 degrees C, 60 min), we found that GSH (10 microM) generated sulfhydryls in the beta 3 subunit. To determine if the requirement was for reducing equivalents or for a redox potential (ratio of GSH to GSSG), aggregation was further studied with the addition of low concentrations of GSSG to the GSH. With a ratio of GSH/GSSG of 5/1, similar to that of blood, the addition of GSSG potentiated the stimulatory effect as compared to GSH alone. This indicates that, for potentiation of aggregation, GSH is not simply reducing disulfide bonds; there is rather a requirement for a certain redox potential. Additional studies performed in the absence of added glutathione showed an increase in sulfhydryl labeling in the beta 3 subunit during platelet activation. Finally, we show that vicinal dithiols of platelet surface proteins are involved in the sulfhydryl-dependent pathways of platelet activation. In summary, these data imply that the redox potential of blood regulates activation of the alpha IIb beta 3 integrin and together with other reports in the literature suggest that disulfide bond cleavage with sulfhydryl generation in beta 3 is involved in activation of this receptor.
authors.affiliation: Department of Medicine, Division of Hematology, University of Texas/Health Science Center at San Antonio, San Antonio, Texas 78229-3900, USA. essex@uthscsa.edu
authors.forename: David W
authors.initials: DW
authors.lastname: Essex
authors.forename: Mengru
authors.initials: M
authors.lastname: Li
mesh_terms: D001152:Arsenicals / Q000378:metabolism / Q000494:pharmacology; D004220:Disulfides / Q000494:pharmacology; D004357:Drug Synergism; D005978:Glutathione / Q000097:blood / Q000494:pharmacology; D019803:Glutathione Disulfide / Q000494:pharmacology; D006801:Humans; D010084:Oxidation-Reduction / Q000187:drug effects; D015539:Platelet Activation / Q000187:drug effects / Q000502:physiology; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D010975:Platelet Aggregation Inhibitors / Q000097:blood / Q000494:pharmacology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014494:Unithiol / Q000494:pharmacology
chemical_list: D001152:Arsenicals; D004220:Disulfides; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D021122:Protein Subunits; D013438:Sulfhydryl Compounds; C029341:oxophenylarsine; D014494:Unithiol; D005978:Glutathione; D019803:Glutathione Disulfide