Expression level dependent changes in the properties of P2X2 receptors.
The currents of P2X(2) receptors expressed in Xenopus oocytes or HEK293 cells show significant cell-to-cell variation in many properties including the rate of desensitization and the magnitude of potentiation by zinc or acidic pH. In this study, we examined whether differences in expression levels underlie this variability. We injected Xenopus oocytes with different concentrations of RNA encoding rat P2X(2) to give a wide range of maximum current amplitudes, and then measured the potentiation of responses to 10 micro M adenosine 5'-triphosphate (ATP) by zinc or acidic pH. Individual oocytes showed potentiation ratios that ranged from 1.4- to 25-fold. Oocytes with small amplitude responses to a saturating concentration of ATP tended to have larger potentiation ratios than oocytes with large amplitude responses. This phenomenon was explained by an inverse correlation between the EC(50) for ATP and the maximum current amplitude, with the EC(50) decreasing from about 37 to 7 micro M as expression level increased. In contrast, the Hill coefficient was not correlated with the maximum current amplitude. Truncated receptors lacking the last 76 amino acids also showed an inverse correlation between the EC(50) and the maximum current amplitude. Thus, the interactions that cause expression-dependent changes in P2X(2) receptor properties must involve domains proximal to position H397.
Clyne
J D
JD
Department of Molecular, Cellular, and Developmental Biology, University of Michigan, 3095 Natural Science Building, 830 N. University Avenue, Ann Arbor, MI 48109-1048, USA.
Brown
T C
TC
Hume
R I
RI
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000187:drug effects*; D006639:Histidine / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D008954:Models, Biological; D016296:Mutagenesis; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000379:methods; D012333:RNA, Messenger / Q000008:administration & dosage; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000235:genetics* / Q000378:metabolism; D058476:Receptors, Purinergic P2X2; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D013329:Structure-Activity Relationship; D014981:Xenopus; D015032:Zinc / Q000494:pharmacology*
C546403:P2rx2 protein, rat; D012333:RNA, Messenger; D018048:Receptors, Purinergic P2; D058476:Receptors, Purinergic P2X2; D011994:Recombinant Proteins; D006639:Histidine; D000255:Adenosine Triphosphate; D015032:Zinc