title: Protein mobility and GABA-induced conformational changes in GABA(A) receptor pore-lining M2 segment.
abstract: Protein movements underlying ligand-gated ion channel activation are poorly understood. Here we used disulfide bond trapping to examine the proximity and mobility of cysteines substituted for aligned GABAA receptor alpha1 and beta1 M2 segment channel-lining residues in resting and activated receptors. With or without GABA, disulfide bonds formed at alpha1N275C/beta1E270C (20') and alpha1S272C/beta1H267C (17'), near the extracellular end, suggesting that this end is more mobile and/or flexible than the rest of the segment. Near the middle of M2, at alpha1T261C/beta1T256C (6'), a disulfide bond formed only in the presence of GABA and locked the channels open. Channel activation must involve an asymmetric rotation of two adjacent subunits toward each other. This would move aligned engineered cysteines on different subunits into proximity and allow disulfide bond formation without blocking conduction. Asymmetric rotation of M2 segments is probably a common gating mechanism in other ligand-gated ion channels.
authors.affiliation: Department of Physiology and Biophysics and Department of Neuroscience, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
authors.forename: J
authors.initials: J
authors.lastname: Horenstein
authors.forename: D A
authors.initials: DA
authors.lastname: Wagner
authors.forename: C
authors.initials: C
authors.lastname: Czajkowski
authors.forename: M H
authors.initials: MH
authors.lastname: Akabas
mesh_terms: D000818:Animals; D004220:Disulfides; D004594:Electrophysiology; D015151:Immunoblotting; D015640:Ion Channel Gating / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011487:Protein Conformation; D051381:Rats; D018079:Receptors, GABA / Q000235:genetics* / Q000378:metabolism*; D014981:Xenopus; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology*
chemical_list: D004220:Disulfides; D009419:Nerve Tissue Proteins; D018079:Receptors, GABA; D005680:gamma-Aminobutyric Acid