Abstract 14937: Protein Network of VASP in Lipid Raft of Platelets
Introduction: VAsodilator-Stimulated Phosphoprotein (VASP) represents a very sensitive marker of platelet responsiveness to ADP receptor (P2Y12) inhibitors. After platelet stimulation, VASP is colocalized with F-actin on Detergent Resistant Membrane (DRM) but the protein network involved is not well known. We aimed to investigate interactive proteins with VASP on DRM of platelets.
Methods: Lipid rafts of human platelets were isolated using Triton X100 1% (w/v) and sucrose gradient. After ultracentrifugation, sucrose fractions were sampled. Lipidomics analysis was performed to identify fractions enriched in cholesterol and sphingomyelin (considered as DRM). These enriched fractions were selected for further proteomic analyses. Post analysis of proteomics results was performed to identify participating protein pathways around VASP.
Results: Sucrose fractions containing platelet DRMs were identified (Fig., left). In these fractions, we identified potential proteins interacting with VASP (Fig, right). Some of the interactions were already confirmed experimentally in Homo Sapiens such as CDC42 and Zyxin (interaction: activation); Vinculin, WAS (Wiskott-Aldrich syndrome effector protein), Ubiquintin and YWHAZ (tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein) (interaction: binding) and GSN (gelsolin) (interaction: inhibition). The association between VASP and others proteins as FYN, FYB, PFN1 (profilin 1), LASP1 (LIM and SH3 protein 1) and SRC were shown experimentally in other species (with their putative homologs). Others proteins in this network are potential candidates for platelet study in order to identify their role in this network and the nature of their relationship with VASP.
Conclusions: We identified proteins associated with VASP in DRM fractions containing rafts of platelet membranes. This could provide further insight into the clinical target of pathways through this molecule.
Author Disclosures: V. Rabani: None. D. Montange: None. N.F. Meneveau: Speakers Bureau; Modest; St Jude Medical, Bayer, Daiichi Sankyo, BMS. S. Davani: None.
- © 2016 by American Heart Association, Inc.