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Specific detection of non‐functional human P2X 7 receptors in HEK293 cells and B‐lymphocytes
Author(s) -
Barden J.A.,
Sluyter R.,
Gu B.J.,
Wiley J.S.
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)00172-8
Subject(s) - hek 293 cells , receptor , transfection , epitope , microbiology and biotechnology , extracellular , mutant , biology , chemistry , antibody , cell culture , biochemistry , immunology , genetics , gene
P2X 7 receptor/channels mediate ATP‐induced apoptosis in a range of cells including lymphocytes. HEK293 cells were transfected with wild‐type human P2X 7 receptor or site‐directed mutant constructs (K193A, K311A and E496A) known to be non‐functional from measurements of barium/ethidium influx in the presence of ATP or 2′,3′‐ O ‐(4‐benzoylbenzoyl)‐ATP. An antibody was designed against an epitope from a loop adjacent to the extracellular ATP site. The epitope was unavailable in cells expressing normal functional surface receptors. Non‐functional surface receptors as well as intracellular receptors selectively bound the antibody. So did B‐lymphocytes from chronic lymphocytic leukemia patients expressing non‐functional (E496A) mutant receptor.