Structure–function relationship of the M2 ion channel of influenza A virus

Jorgen A. Mould, Kevin Shuck, Jason E. Drury, Stephan M. Frings, U. Benjamin Kaupp, Andrew Pekosz, Robert A. Lamb, Lawrence H. Pinto

Research output: Contribution to journalArticle

Abstract

The use of oligomers comprised of amantadine-sensitive and -resistant forms demonstrated that the active oligomeric state of the channel is a tetramer. Cysteine scanning mutagenesis followed by evaluation of the ability of sulfhydryl-specific reagents to inhibit the channel demonstrated that each monomer is a coiled coil and that the pore-lining residues are Val-27, Ala-30, Gly-34, His-37 and Trp-41. Under oxidizing conditions, mutant proteins containing cysteine at or close to these residues form dimers, but do so less readily at low pH for residues near #41, supporting the notion that a pH-induced conformational change occurs in this region of the protein. A functional change in state was detected by comparing the efflux of H+ from acid-loaded cells expressing M2 protein with those treated with the protonophore FCCP: the efflux was high for FCCP-treated cells, but not M2-expressing cells when pHout was elevated. The current of the wild-type protein is carried by H+ and is increased by low pHout, but replacement of the His-37 results in pH-independent currents of other ions as well, suggesting that the selectivity and activation of the channel might result from the action of His-37. The C-terminus is needed for sustained function of the channel, as truncation mutants are abnormal and that they pass H+ for only a short time.

Original languageEnglish (US)
Pages (from-to)389-396
Number of pages8
JournalInternational Congress Series
Volume1219
Issue numberC
DOIs
StatePublished - Oct 1 2001
Externally publishedYes

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Influenza A virus
Ion Channels
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Cysteine
Amantadine
Sulfhydryl Reagents
Proteins
Mutant Proteins
Mutagenesis
Ions

Keywords

  • Amantadine
  • Ion channels

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Mould, J. A., Shuck, K., Drury, J. E., Frings, S. M., Kaupp, U. B., Pekosz, A., ... Pinto, L. H. (2001). Structure–function relationship of the M2 ion channel of influenza A virus. International Congress Series, 1219(C), 389-396. https://doi.org/10.1016/S0531-5131(01)00641-0

Structure–function relationship of the M2 ion channel of influenza A virus. / Mould, Jorgen A.; Shuck, Kevin; Drury, Jason E.; Frings, Stephan M.; Kaupp, U. Benjamin; Pekosz, Andrew; Lamb, Robert A.; Pinto, Lawrence H.

In: International Congress Series, Vol. 1219, No. C, 01.10.2001, p. 389-396.

Research output: Contribution to journalArticle

Mould, JA, Shuck, K, Drury, JE, Frings, SM, Kaupp, UB, Pekosz, A, Lamb, RA & Pinto, LH 2001, 'Structure–function relationship of the M2 ion channel of influenza A virus', International Congress Series, vol. 1219, no. C, pp. 389-396. https://doi.org/10.1016/S0531-5131(01)00641-0
Mould, Jorgen A. ; Shuck, Kevin ; Drury, Jason E. ; Frings, Stephan M. ; Kaupp, U. Benjamin ; Pekosz, Andrew ; Lamb, Robert A. ; Pinto, Lawrence H. / Structure–function relationship of the M2 ion channel of influenza A virus. In: International Congress Series. 2001 ; Vol. 1219, No. C. pp. 389-396.
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