The unanticipated complexity of the selectivity-filter glutamates of nicotinic receptors.
In ion channels, 'rings' of ionized side chains that decorate the walls of the permeation pathway often lower the energetic barrier to ion conduction. Using single-channel electrophysiological recordings, we studied the poorly understood ring of four glutamates (and one glutamine) that dominates this catalytic effect in the muscle nicotinic acetylcholine receptor ('the intermediate ring of charge'). We show that all four wild-type glutamate side chains are deprotonated in the range of 6.0-9.0 pH, that only two of them contribute to the size of the single-channel current, that these side chains must be able to adopt alternate conformations that either allow or prevent their negative charges from increasing the rate of cation conduction and that the location of these glutamate side chains squarely at one of the ends of the transmembrane pore is critical for their largely unshifted pK(a) values and for the unanticipated impact of their conformational flexibility on cation permeation.
Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Gisela D
GD
Cymes
Claudio
C
Grosman
D000328:Adult; D000595:Amino Acid Sequence; D002384:Catalysis; D002412:Cations / Q000378:metabolism; D017666:Deuterium Oxide / Q000378:metabolism; D004563:Electrochemistry; D005971:Glutamates / Q000737:chemistry*; D005973:Glutamine / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology; D008968:Molecular Conformation; D018482:Muscle, Skeletal / Q000201:enzymology; D018408:Patch-Clamp Techniques; D016062:Porosity; D011978:Receptors, Nicotinic / Q000737:chemistry*
D002412:Cations; D005971:Glutamates; D007473:Ion Channels; D011978:Receptors, Nicotinic; D005973:Glutamine; D017666:Deuterium Oxide