title: Vicinal thiols are required for activation of the αIIbβ3 platelet integrin.
abstract: BACKGROUND
  Closely spaced thiols in proteins that interconvert between the dithiol form and disulfide bonds are called vicinal thiols. These thiols provide a mechanism to regulate protein function. We previously found that thiols in both αIIb and β3 of the αIIbβ3 fibrinogen receptor were required for platelet aggregation.
  
  
  RESULTS
  Using p-chloromercuribenzene sulfonate (pCMBS) we provide evidence that surface thiols in αIIbβ3 are exposed during platelet activation. Phenylarsine oxide (PAO), a reagent that binds vicinal thiols, inhibits platelet aggregation and labeling of sulfhydryls in both αIIb and β3. For the aggregation and labeling studies, binding of PAO to vicinal thiols was confirmed by reversal of PAO binding with the dithiol reagent 2,3-Dimercapto-1-propanesulfonic acid (DMPS). In contrast, the monothiol β-mercaptoethanol did not reverse the effects of PAO. Additionally, PAO did not inhibit sulfhydryl labeling of the monothiol protein albumin, confirming the specificity of PAO for vicinal thiols in αIIbβ3. As vicinal thiols represent redox sensitive sites that can be regulated by reducing equivalents from the extracellular or cytoplasmic environment, they are likely to be important in regulating activation of αIIbβ3. Additionally, when the labeled integrin was passed though a lectin column containing wheat germ agglutinin and lentil lectin a substantial amount of non-labeled αIIbβ3 eluted separately from the labeled receptor. This suggests that two populations of integrin exist on platelets that can be distinguished by thiol labeling.
  
  
  CONCLUSIONS
  A vicinal thiol-containing population of αIIbβ3 provides redox sensitive sites for regulation of αIIbβ3.
authors.affiliation: Division of Hematology, Department of Medicine, The University of Texas Health Science Center, San Antonio, TX , USA.
authors.forename: N
authors.initials: N
authors.lastname: Manickam
authors.forename: S S
authors.initials: SS
authors.lastname: Ahmad
authors.forename: D W
authors.initials: DW
authors.lastname: Essex
mesh_terms: D001152:Arsenicals / Q000494:pharmacology; D001792:Blood Platelets / Q000166:cytology; D002478:Cells, Cultured; D006801:Humans; D039421:Integrin alpha2 / Q000737:chemistry / Q000378:metabolism; D039661:Integrin beta3 / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D010084:Oxidation-Reduction; D010974:Platelet Aggregation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism* / Q000502:physiology
chemical_list: D001152:Arsenicals; D039421:Integrin alpha2; D039661:Integrin beta3; D008565:Membrane Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D013438:Sulfhydryl Compounds; C029341:oxophenylarsine