A unified view of the role of electrostatic interactions in modulating the gating of Cys loop receptors.
In the Cys loop superfamily of ligand-gated ion channels, a global conformational change, initiated by agonist binding, results in channel opening and the passage of ions across the cell membrane. The detailed mechanism of channel gating is a subject that has lent itself to both structural and electrophysiological studies. Here we defined a gating interface that incorporates elements from the ligand binding domain and transmembrane domain previously reported as integral to proper channel gating. An overall analysis of charged residues within the gating interface across the entire superfamily showed a conserved charging pattern, although no specific interacting ion pairs were conserved. We utilized a combination of conventional mutagenesis and the high precision methodology of unnatural amino acid incorporation to study extensively the gating interface of the mouse muscle nicotinic acetylcholine receptor. We found that charge reversal, charge neutralization, and charge introduction at the gating interface are often well tolerated. Furthermore, based on our data and a reexamination of previously reported data on gamma-aminobutyric acid, type A, and glycine receptors, we concluded that the overall charging pattern of the gating interface, and not any specific pairwise electrostatic interactions, controls the gating process in the Cys loop superfamily.
Xiu
Xinan
X
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Hanek
Ariele P
AP
Wang
Jinti
J
Lester
Henry A
HA
Dougherty
Dennis A
DA
D000109:Acetylcholine / Q000737:chemistry; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001671:Biochemistry / Q000379:methods; D015153:Blotting, Western; D002038:Bungarotoxins / Q000737:chemistry; D002412:Cations; D002462:Cell Membrane / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry*; D003553:Cystine / Q000737:chemistry; D030562:Databases, Protein; D004305:Dose-Response Relationship, Drug; D004566:Electrodes; D004594:Electrophysiology; D005998:Glycine / Q000737:chemistry; D007477:Ions; D007700:Kinetics; D008024:Ligands; D051379:Mice; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D055672:Static Electricity; D014101:Torpedo; D014982:Xenopus laevis; D005680:gamma-Aminobutyric Acid / Q000737:chemistry
D000596:Amino Acids; D002038:Bungarotoxins; D002412:Cations; D007477:Ions; D008024:Ligands; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D003553:Cystine; D005680:gamma-Aminobutyric Acid; D003545:Cysteine; D000109:Acetylcholine; D005998:Glycine