Synthesis and characterization of a cellular membrane affinity chromatography column containing histamine 1 and P2Y(1) receptors: a multiple G-protein coupled receptor column.
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.
Moaddel
Ruin
R
National Institute on Aging, National Institutes of Health, Gerontology Research Center, 5600 Nathan Shock Drive, Baltimore, MD 21224-6825, United States. moaddelru@mail.nih.gov
Musyimi
Harrison K
HK
Sanghvi
Mitesh
M
Bashore
Charlene
C
Frazier
Chester R
CR
Khadeer
Mohammad
M
Bhatia
Prateek
P
Wainer
Irving W
IW
D001254:Astrocytoma / Q000473:pathology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000295:instrumentation*; D006801:Humans; D007700:Kinetics; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011968:Receptors, Histamine / Q000378:metabolism*; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D014162:Transfection
D043562:Receptors, G-Protein-Coupled; D011968:Receptors, Histamine; D018048:Receptors, Purinergic P2