Specific detection of non‐functional human P2X7 receptors in HEK293 cells and B‐lymphocytes

2003/02/22 by Julian A. Barden, J.A. Barden, R. Sluyter +5 · 12 citations
Biochemistry, Genetics and Molecular Biology · Immunology and Microbiology · Health Professions · #Adenosine and Purinergic Signaling #Immune Cell Function and Interaction #Adolescent and Pediatric Healthcare

paper · pdf · doi:10.1016/s0014-5793(03)00172-8

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.

Citations

• Point mutations confer loss of ATP‐induced human P2X7 receptor function

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