Abstract
P2X receptors are a family of ion channels gated by extracellular ATP. Each member of the family can form functional homomeric channels, but only P2X2 and P2X3 have been shown to combine to form a unique heteromeric channel. Data from in situhybridization studies suggest that P2X1 and P2X5 may also co-assemble. In this study, we tested this hypothesis by expressing recombinant P2X1 and P2X5 receptor subunits either individually or together in human embryonic kidney 293 cells. In cells expressing the homomeric P2X1 receptor, 30 μm α,β-methylene ATP (α,β-me-ATP) evoked robust currents that completely desensitized in less than 1 sec, whereas α,β-me-ATP failed to evoke current in cells expressing the homomeric P2X5 receptor. By contrast, α,β-me-ATP evoked biphasic currents with a pronounced nondesensitizing plateau phase in cells that co-expressed both subunits. Further, the EC50 for α,β-me-ATP was greater in cells expressing both P2X1 and P2X5 than in cells expressing P2X1 alone (5 and 1.6 μm, respectively). Heteromeric assembly was confirmed using a co-immunoprecipitation assay of epitope-tagged P2X1 and P2X5 subunits. In summary, this study provides biochemical and functional evidence of a novel channel formed by P2X subunit heteropolymerization. This finding suggests that heteromeric subunit assembly constitutes an important mechanism for generating functional diversity of ATP-mediated responses.
Footnotes
- Received July 31, 1998.
- Accepted August 25, 1998.
-
Send reprint requests to: Dr. Mark M. Voigt, Department of Pharmacological and Physiological Science, St. Louis University School of Medicine, 1402 South Grand Blvd. Saint Louis, MO 63104. E-mail:voigtm{at}slu.edu
-
Supported by National Institutes of Health Grants HL56236 (T.M.E.) and NS35534 (M.M.V.) and an American Heart Association–Missouri Affiliate predoctoral fellowship (W.R.H). G.E.T. and W.R.H. contributed equally to this work.
- The American Society for Pharmacology and Experimental Therapeutics
MolPharm articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|