Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the α subunit

Myles Akabas, Christine Kaufmann, Patrick Archdeacon, Arthur Karlin

Research output: Contribution to journalArticle

350 Citations (Scopus)

Abstract

Each residue in and flanking the M2 membrane-spanning segment of the α subunit, from Glu-241 to Glu-262, was mutated to cysteine, and the mutant subunits were expressed together with wild-type β, γ, and δ subunits in Xenopus oocytes. Cysteines substituted for Glu-262, Leu-258, Val-255, Ser-252, Leu-251, Leu-250, Ser-248, Leu-245, Thr-244, and Glu-241 reacted with the positively charged, hydrophilic, sulfhydryl-specific reagent methanethiosulfonate ethylammonium (MTSEA), added extracellularly. These 10 residues, therefore, are exposed in the channel lumen. The pattern of exposure is compatible with an α helix, interrupted by an extended structure from Leu-250 to Ser-252. Acetylcholine caused subtle changes in the accessibilities of some of the engineered cysteines. Since all 10 residues are accessible to MTSEA in the closed state of the channel, the channel gate is at least as cytoplasmic as Glu-241, the most cytoplasmic of the residues tested.

Original languageEnglish (US)
Pages (from-to)919-927
Number of pages9
JournalNeuron
Volume13
Issue number4
DOIs
StatePublished - 1994
Externally publishedYes

Fingerprint

Cholinergic Receptors
Cysteine
Sulfhydryl Reagents
Xenopus
Acetylcholine
Oocytes
Membranes
methanethiosulfonate ethylammonium

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the α subunit. / Akabas, Myles; Kaufmann, Christine; Archdeacon, Patrick; Karlin, Arthur.

In: Neuron, Vol. 13, No. 4, 1994, p. 919-927.

Research output: Contribution to journalArticle

Akabas, Myles ; Kaufmann, Christine ; Archdeacon, Patrick ; Karlin, Arthur. / Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the α subunit. In: Neuron. 1994 ; Vol. 13, No. 4. pp. 919-927.
@article{0c58c0278bf742ca98f0542ce9f0e294,
title = "Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the α subunit",
abstract = "Each residue in and flanking the M2 membrane-spanning segment of the α subunit, from Glu-241 to Glu-262, was mutated to cysteine, and the mutant subunits were expressed together with wild-type β, γ, and δ subunits in Xenopus oocytes. Cysteines substituted for Glu-262, Leu-258, Val-255, Ser-252, Leu-251, Leu-250, Ser-248, Leu-245, Thr-244, and Glu-241 reacted with the positively charged, hydrophilic, sulfhydryl-specific reagent methanethiosulfonate ethylammonium (MTSEA), added extracellularly. These 10 residues, therefore, are exposed in the channel lumen. The pattern of exposure is compatible with an α helix, interrupted by an extended structure from Leu-250 to Ser-252. Acetylcholine caused subtle changes in the accessibilities of some of the engineered cysteines. Since all 10 residues are accessible to MTSEA in the closed state of the channel, the channel gate is at least as cytoplasmic as Glu-241, the most cytoplasmic of the residues tested.",
author = "Myles Akabas and Christine Kaufmann and Patrick Archdeacon and Arthur Karlin",
year = "1994",
doi = "10.1016/0896-6273(94)90257-7",
language = "English (US)",
volume = "13",
pages = "919--927",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "4",

}

TY - JOUR

T1 - Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the α subunit

AU - Akabas, Myles

AU - Kaufmann, Christine

AU - Archdeacon, Patrick

AU - Karlin, Arthur

PY - 1994

Y1 - 1994

N2 - Each residue in and flanking the M2 membrane-spanning segment of the α subunit, from Glu-241 to Glu-262, was mutated to cysteine, and the mutant subunits were expressed together with wild-type β, γ, and δ subunits in Xenopus oocytes. Cysteines substituted for Glu-262, Leu-258, Val-255, Ser-252, Leu-251, Leu-250, Ser-248, Leu-245, Thr-244, and Glu-241 reacted with the positively charged, hydrophilic, sulfhydryl-specific reagent methanethiosulfonate ethylammonium (MTSEA), added extracellularly. These 10 residues, therefore, are exposed in the channel lumen. The pattern of exposure is compatible with an α helix, interrupted by an extended structure from Leu-250 to Ser-252. Acetylcholine caused subtle changes in the accessibilities of some of the engineered cysteines. Since all 10 residues are accessible to MTSEA in the closed state of the channel, the channel gate is at least as cytoplasmic as Glu-241, the most cytoplasmic of the residues tested.

AB - Each residue in and flanking the M2 membrane-spanning segment of the α subunit, from Glu-241 to Glu-262, was mutated to cysteine, and the mutant subunits were expressed together with wild-type β, γ, and δ subunits in Xenopus oocytes. Cysteines substituted for Glu-262, Leu-258, Val-255, Ser-252, Leu-251, Leu-250, Ser-248, Leu-245, Thr-244, and Glu-241 reacted with the positively charged, hydrophilic, sulfhydryl-specific reagent methanethiosulfonate ethylammonium (MTSEA), added extracellularly. These 10 residues, therefore, are exposed in the channel lumen. The pattern of exposure is compatible with an α helix, interrupted by an extended structure from Leu-250 to Ser-252. Acetylcholine caused subtle changes in the accessibilities of some of the engineered cysteines. Since all 10 residues are accessible to MTSEA in the closed state of the channel, the channel gate is at least as cytoplasmic as Glu-241, the most cytoplasmic of the residues tested.

UR - http://www.scopus.com/inward/record.url?scp=0027987062&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027987062&partnerID=8YFLogxK

U2 - 10.1016/0896-6273(94)90257-7

DO - 10.1016/0896-6273(94)90257-7

M3 - Article

VL - 13

SP - 919

EP - 927

JO - Neuron

JF - Neuron

SN - 0896-6273

IS - 4

ER -