Threonine in the selectivity filter of the acetylcholine receptor channel.
The acetylcholine receptor (AChR) is a cation selective channel whose biophysical properties as well as its molecular composition are fairly well characterized. Previous studies on the rat muscle alpha-subunit indicate that a threonine residue located near the cytoplasmic side of the M2 segment is a determinant of ion flow. We have studied the role of this threonine in ionic selectivity by measuring conductance sequences for monovalent alkali cations and bionic reversal potentials of the wild type (alpha beta gamma delta channel) and two mutant channels in which this threonine was replaced by either valine (alpha T264V) or glycine (alpha T264G). For the wild type channel we found the selectivity sequence Rb greater than Cs greater than K greater than Na. The alpha T264V mutant channel had the sequence Rb greater than K greater than Cs greater than Na. The alpha T264G mutant channel on the other hand had the same selectivity sequence as the wild type, but larger permeability ratios Px/PNa for the larger cations. Conductance concentration curves indicate that the effect of both mutations is to change both the maximum conductance as well as the apparent binding constant of the ions to the channel. A difference in Mg2+ sensitivity between wild-type and mutant channels, which is a consequence of the differences in ion binding, was also found. The present results suggest that alpha T264 form part of the selectivity filter of the AChR channel were large ions are selected according to their dehydrated size.
Villarroel
A
A
Abteilung Zellphysiologie, Max-Planck-Institut für Medizinische, Heidelberg, Germany.
Sakmann
B
B
D000595:Amino Acid Sequence; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D004553:Electric Conductivity; D005260:Female; D007473:Ion Channels / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D010539:Permeability; D011487:Protein Conformation; D051381:Rats; D011950:Receptors, Cholinergic / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics; D013912:Threonine / Q000737:chemistry*; D014981:Xenopus
D007473:Ion Channels; D011950:Receptors, Cholinergic; D013912:Threonine; D008274:Magnesium