Association of Membrane/Lipid Rafts With the Platelet Cytoskeleton and the Caveolin PY14: Participation in the Adhesion Process

Abstract

Platelets are the most prominent elements of blood tissue involved in hemostasis at sites of blood vessel injury. Platelet cytoskeleton is responsible for their shape modifications observed during activation and adhesion to the substratum; therefore the interactions between cytoskeleton and plasma membrane are critical to modulate blood platelet functions. Several cytoskeletal components and binding partners, as well as enzymes that regulate the cytoskeleton, localize to membrane/lipid rafts (MLR) and regulate lateral diffusion of membrane proteins and lipids. Resting, thrombin-activated, and adherent human platelets were processed for biochemical studies including western-blot and immunprecipitation assays and confocal analysis were performed to characterize the interaction of MLR with the main cytoskeleton elements and β-dystroglycan as well as with the association of caveolin-1 PY14 with focal adhesion proteins. We transfected a megakaryoblast cell line (Meg-01) to deplete β-dystroglycan, subsequent to their differentiation to the platelet progenitors. Our data showed a direct interaction of the MLR with cytoskeleton to regulate platelet shape, while an association of caveolin-1 PY14 with vinculin is needed to establish focal adhesions, which are modulated for β-dystroglycan. In conclusion, caveolin-1 PY14 in association with platelet cytoskeleton participate in focal adhesions dynamics.

Platelet membrane lipid rafts protein composition varies following GPVI and CLEC-2 receptors activation

“…Most proteins identified in our LC-MS/MS analysis are related to cell signalling and cytoskeleton-remodelling events necessary for platelet function. This observation strengthens the idea of the requirement of lipid rafts for signal transduction as well as their role in the cross talk between membrane proteins and the cytoskeleton [8,29,30].…”

Pharmaceutical Technology

“…Biological material. Blood was donated by Banco de Sangre, Hospital Juárez de México, and platelets from the blood were obtained in the Laboratorio de Hematobiología, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional (IPN), according to a standardized method as previously described by Cerecedo et al (13). Platelet-rich plasma was collected by centrifugation of whole blood at 400 g for 10 minutes at room temperature with an equal volume of citrate anticoagulant.…”

Dystrophin–Glycoprotein Complex in Blood Cells

“…In this regard, cholesterol demonstrated to be essential to modulate platelet cytoskeleton reorganization, while the association of caveolin-1 PY14 with intermediate ilaments, as well as with focal adhesion proteins via vinculin, was a determinant in adhered platelets, where β-Dg participation was a key scafold component for caveolin-1 and FAK [106].…”