Binding of a thrombin receptor tethered ligand analogue to human platelet thrombin receptor

被引:57
作者
Ahn, HS
Foster, C
Boykow, G
Arik, L
SmithTorhan, A
Hesk, D
Chatterjee, M
机构
[1] Schering-Plough Research Institute, Kenilworth
[2] Schering-Plough Research Institute, Kenilworth
关键词
D O I
10.1124/mol.51.2.350
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
A thrombin receptor-radioligand binding assay was developed using [H-3]A(pF-F)R(ChA)(hR)Y-NH2 ([H-3]haTRAP), a high affinity thrombin receptor-activating peptide (TRAP), and human platelet membranes. Scatchard analysis of saturation binding data indicated that [H-3]haTRAP bound to platelet membranes with a K-d of 15 nM and a B-max of 5.2 pmol/mg of protein. The binding was reduced by GPPNHP, a nonmetabolizable GTP analogue. Various TRAPs and a TRAP antagonist, but not other receptor agonists, displaced [H-3]haTRAP from the binding sites. SFLLRN-NH2, a thrombin receptor-tethered ligand analogue, and [H-3]haTRAP exhibited competitive binding for the same binding sites. The relative affinity of these peptides for the binding site paralleled their EC(50) or IC50 values for platelet aggregation. These data indicate that [H-3]haTRAP binds specifically and saturably to the functioning G protein-linked thrombin (tethered ligand) receptor in human platelet membranes.
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页码:350 / 356
页数:7
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