The adherence of platelets to adsorbed albumin by receptor-mediated recognition of binding sites exposed by adsorption-induced unfolding.
Although albumin (Alb) is the most abundant plasma protein, it is considered to be non-adhesive to platelets, as it lacks any known amino acid sequences for binding platelet receptors. Recent studies have suggested that platelets adhere to adsorbed Alb by mechanisms linked to its conformational state. To definitively address this issue we used circular dichroism (CD) spectropolarimetry to characterize the conformation of Alb adsorbed on a broad range of surface chemistries from a wide range of Alb solution concentrations, with platelet adhesion examined using a lactate dehydrogenase (LDH) assay and scanning electron microscopy (SEM). Our results prove that platelets bind to adsorbed Alb through receptor-mediated processes, with binding sites in Alb exposed and/or formed by adsorption-induced protein unfolding. Most importantly, beyond a critical degree of unfolding, the platelet adhesion levels correlated strongly with the adsorption-induced unfolding in Alb. The blockage of Arg-Gly-Asp (RGD) specific platelet receptors using an Arg-Gly-Asp-Ser (RGDS) peptide led to significant inhibition of platelet adhesion to adsorbed Alb, with the extent of inhibition and morphology of adherent platelets being similar for both Alb and Fg. Chemical neutralization of arginine (Arg) residues in the adsorbed Alb layer inhibited platelet-Alb interactions significantly, indicating that Arg residues play a prominent role in mediating platelet adhesion to Alb. These results provide deeper insight into the molecular mechanisms that mediate the interactions of platelets with adsorbed proteins, and how to control these interactions to improve the blood compatibility of biomaterials for cardiovascular applications.
Sivaraman
Balakrishnan
B
Department of Bioengineering, 501 Rhodes Engineering Center, Clemson University, Clemson, SC 29634, USA.
Latour
Robert A
RA
D000327:Adsorption; D001665:Binding Sites; D001792:Blood Platelets / Q000737:chemistry* / Q000378:metabolism*; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D006801:Humans; D008422:Materials Testing; D010973:Platelet Adhesiveness / Q000502:physiology*; D011485:Protein Binding; D011489:Protein Denaturation; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism*
D020099:Coated Materials, Biocompatible; D012709:Serum Albumin