Effects of pH on acetylcholine receptor function.
We have examined the effects of changing extracellular pH on the function of nicotinic acetylcholine receptors from Torpedo californica using ion flux and electrophysiological methods. Agonist-induced cation efflux from vesicles containing purified, reconstituted receptors showed a monotonic dependence on external hydrogen ion concentration with maximal fluxes at alkaline pH and no agonist-induced efflux at pH's less than approximately 5. A similar pH dependence was measured for the peak agonist-activated membrane currents measured in microelectrode voltage-clamped Xenopus oocytes induced to express Torpedo receptor through mRNA injection. Half-maximal inhibition occurred at a similar pH in both systems, in the range of pH 6.5-7.0. Single-channel currents from Torpedo ACh receptors measured in patch-clamp recordings were also reduced in amplitude at acid pH with an apparent pKa for block of less than 5. Measurements of channel kinetics had a more complicated dependence on pH. The mean channel open time determined from patch-clamp measurements was maximal at neutral pH and decreased at both acid and alkaline pH's. Thus, both channel permeability properties and channel gating properties are affected by the extracellular pH.
Palma
A
A
Department of Biochemistry, University of California, Davis 95616.
Li
L
L
Chen
X J
XJ
Pappone
P
P
McNamee
M
M
D000818:Animals; D002038:Bungarotoxins / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D002463:Cell Membrane Permeability; D004553:Electric Conductivity; D004594:Electrophysiology; D006863:Hydrogen-Ion Concentration; D015640:Ion Channel Gating; D008081:Liposomes; D008564:Membrane Potentials; D008839:Microelectrodes; D009865:Oocytes / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D011978:Receptors, Nicotinic / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014101:Torpedo; D014982:Xenopus laevis
D002038:Bungarotoxins; D008081:Liposomes; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D002217:Carbachol