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Molecular characterization of human platelet glycoproteins IIIa and IIb and the subunits of the latter
Authors:P Usobiaga  J J Calvete  J L Saíz  M T Eirín  J González-Rodríguez
Institution:(1) Instituto de Química Física, C.S.I.C., Serrano, 119, E-28006 Madrid, Spain
Abstract:Sedimentation equilibrium and low-angle laser-light scattering were used to determine the molar mass of the glycoprotein moieties in the complexes of sodium dodecyl sulphate with the human platelet membrane glycoproteins IIb (GPIIb), IIIa (GPIIIa), and the agr (GPIIbagr) and beta (GPIIbbeta) subunits of GPIIb. The values obtained by both procedures, except those for GPIIb, agree within experimental error with those calculated from their chemical composition: GPIIbagr (114,000 g mol-1), GPIIbbeta (22,200 g mol-1), and GPIIIa (91,500 g mol-1). The molar mass of GPIIb determined by light scattering (142,000 g mol-1) and sedimentation equilibrium at different solvent densities (134,000 g mol-1) also agree, within experimental error, with the values calculated either from its chemical composition (136,500 g mol-1) or from the sum of the molar masses of its subunits. However the molar mass determined by sedimentation equilibrium at constant solvent density, is consistently underestimated (116,000 g mol-1).High-performance size-exclusion chromatography in sodium dodecyl sulphate solutions overestimates the molar mass of these glycoproteins and their Stokes radii, and therefore the maximal frictional ratios derived from them.Abbreviations GPIIb glycoprotein IIb - GPIIIa glycoprotein IIIa - GPIIbagr and GPIIbbeta agr and beta subunits of GPIIb, respectively - CM-GPIIbagr CM-GPIIbbeta, and CM-GPIIIa, totally reduced and carboxymethylated forms of GPIIbagr, GPIIbbeta, and GPIIIa, respectively - SDS sodium dodecyl sulphate - eosin-ITC eosin-5-isothiocyanate
Keywords:Molar mass  size  shape  platelet glycoproteins  sedimentation equilibrium  laser-light scattering
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