Amino acid residues lining the chloride channel of the cystic fibrosis transmembrane conductance regulator

Myles Akabas, Christine Kaufmann, Tiffani A. Cook, Patrick Archdeacon

Research output: Contribution to journalArticle

134 Citations (Scopus)

Abstract

The cystic fibrosis transmembrane conductance regulator forms a chloride channel that is regulated by phosphorylation and intracellular ATP levels. The structure of the channel-forming domains is undetermined. To identify the residues lining this channel we substituted cysteine, one at a time, for 9 consecutive residues (91-99) in the M1 membrane-spanning segment. The cysteine substitution mutants were expressed in Xenopus oocytes. We determined the accessibility of the engineered cysteine to charged, sulfhydryl-specific methanethiosulfonate reagents added extracellularly. We assume that, among residues in membrane-spanning segments, only those lining the channel will be accessible to react with these hydrophilic reagents and that such a reaction would irreversibly alter conduction through the channel. Only the cysteines substituted for Gly-91, Lys-95, and Gln-98 were accessible to the reagents. We conclude that these residues are in the channel lining. The periodicity of these residues is consistent with an α-helical secondary structure.

Original languageEnglish (US)
Pages (from-to)14865-14868
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number21
StatePublished - May 27 1994
Externally publishedYes

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Cystic Fibrosis Transmembrane Conductance Regulator
Chloride Channels
Linings
Cysteine
Amino Acids
Membranes
Phosphorylation
Periodicity
Xenopus
Oocytes
Substitution reactions
Adenosine Triphosphate

ASJC Scopus subject areas

  • Biochemistry

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Amino acid residues lining the chloride channel of the cystic fibrosis transmembrane conductance regulator. / Akabas, Myles; Kaufmann, Christine; Cook, Tiffani A.; Archdeacon, Patrick.

In: Journal of Biological Chemistry, Vol. 269, No. 21, 27.05.1994, p. 14865-14868.

Research output: Contribution to journalArticle

Akabas, Myles ; Kaufmann, Christine ; Cook, Tiffani A. ; Archdeacon, Patrick. / Amino acid residues lining the chloride channel of the cystic fibrosis transmembrane conductance regulator. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 21. pp. 14865-14868.
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