alexa Specific detection of non-functional human P2X(7) receptors in HEK293 cells and B-lymphocytes.


Journal of Clinical & Cellular Immunology

Author(s): Barden JA, Sluyter R, Gu BJ, Wiley JS

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Abstract 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.
This article was published in FEBS Lett and referenced in Journal of Clinical & Cellular Immunology

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