Synthesis and characterization of a cellular membrane affinity chromatography column containing histamine 1 and P2Y(1) receptors: a multiple G-protein coupled receptor column

含组胺1和P2Y(1)受体的细胞膜亲和层析柱的合成与表征:多G蛋白偶联受体柱

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作者:Ruin Moaddel, Harrison K Musyimi, Mitesh Sanghvi, Charlene Bashore, Chester R Frazier, Mohammad Khadeer, Prateek Bhatia, Irving W Wainer

Abstract

A cellular membrane affinity chromatography (CMAC) column has been created using cellular membrane fragments from a 1321N1 cell line stably transfected with the P2Y(1) receptor. The CMAC(1321N1(P2Y1)) column contained functional P2Y(1) and histamine 1 receptors, which independently bound receptor-specific ligands. The data obtained with the CMAC(1321N1(P2Y1)) column demonstrate that multiple-G-protein coupled receptor (GPCR) columns can be developed and used to probe interactions with the immobilized receptors and that endogenously expressed GPCRs can be used to create CMAC columns. The results also establish that the histamine 1 receptor can be immobilized with retention of ligand-specific binding.

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