Differential modulation of beta2 and beta4 subunits of human neuronal nicotinic acetylcholine receptors by acidification.
We have shown previously that acidification increases the affinity of agonists to rat alpha3beta4 nicotinic acetylcholine receptors (nAChR) and accelerates both the activation and decay kinetics of agonist-induced currents recorded from human embryonic kidney 293 cells stably expressing the receptor (Abdrakhmanova et al., 2002b). Here, we report on experiments examining the effect of rapid acidification on four different subtypes (alpha3beta4alpha5, alpha4beta2, alpha3beta2, and alpha3beta2alpha5) of human neuronal nAChRs stably expressed in tsA201 cells using a piezoelectric device for rapid (<5 ms) solution application. Application of ACh, at its EC(50) concentration for each nAChR subtype, at pH values 7.4 and 6.0, showed that acidification, similarly to that reported for rat alpha3beta4 acetylcholine receptors (AChRs), increased the amplitude and accelerated the activation and decay kinetics of the currents in human alpha3beta4alpha5 AChRs by increasing their affinity to the agonist. In sharp contrast, acidification reduced the amplitude but accelerated the decay kinetics of the current in all human beta2-containing nAChR subtypes (alpha3beta2, alpha3alpha5beta2, alpha4beta2) examined in this study. Brief application of ACh at saturating concentration (1 mM) on alpha3beta4alpha5 AChRs induced a "rebound current" upon rapid washout of the agonist at pH 7.4, but no "rebound current" was observed in alpha3beta2 AChRs. Surprisingly, acidification, pH 6.0, applied only during the agonist pulse also accelerated the decay kinetics of the "rebound current". Our data provide evidence for the specificity of proton-induced modulation of neuronal nAChRs based on their beta subunit composition. Furthermore, in alpha3beta4alpha5 AChR, we find that protonation effects may persist, after washout of acidic solutions, consistent with proton-induced conformational changes of the receptor.
Abdrakhmanova
Galya
G
Department of Pharmacology, Georgetown University School of Medicine, 4000 Reservoir Road Building D, Washington DC 20007, USA.
Cleemann
Lars
L
Lindstrom
Jon
J
Morad
Martin
M
D002478:Cells, Cultured; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009474:Neurons / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000378:metabolism*; D011522:Protons; D011978:Receptors, Nicotinic / Q000378:metabolism*; D014162:Transfection
D021122:Protein Subunits; D011522:Protons; D011978:Receptors, Nicotinic; C403530:nicotinic receptor alpha3beta4