Identification of the functional core of the influenza A virus A/M2 proton-selective ion channel

Chunlong Ma, Alexei L. Polishchuk, Yuki Ohigashi, Amanda L. Stouffer, Arne Schön, Emma Magavern, Xianghong Jing, James D. Lear, Ernesto Freire, Robert A. Lamb, William F. DeGrado, Lawrence H. Pinto

Research output: Contribution to journalArticlepeer-review

121 Scopus citations

Abstract

The influenza A virus M2 protein (A/M2) is a homotetrameric pH-activated proton transporter/channel that mediates acidification of the interior of endosomally encapsulated virus. This 97-residue protein has a single transmembrane (TM) helix, which associates to form homotetramers that bind the anti-influenza drug amantadine. However, the minimal fragment required for assembly and proton transport in cellular membranes has not been defined. Therefore, the conductance properties of truncation mutants expressed in Xenopus oocytes were examined. A short fragment spanning residues 21-61, M2(21-61), was inserted into the cytoplasmic membrane and had specific, amantadine-sensitive proton transport activity indistinguishable from that of full-length A/M2; an epitope-tagged version of an even shorter fragment, M2(21-51)-FLAG, had specific activity within a factor of 2 of the full-length protein. Furthermore, synthetic fragments including a peptide spanning residues 22-46 were found to transport protons into liposomes in an amantadine-sensitive manner. In addition, the functionally important His-37 residue pKa values are highly perturbed in the tetrameric form of the protein, a property conserved in the TM peptide and full-length A/M2 in both micelles and bilayers. These data demonstrate that the determinants for folding, drug binding, and proton translocation are packaged in a remarkably small peptide that can now be studied with confidence.

Original languageEnglish (US)
Pages (from-to)12283-12288
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number30
DOIs
StatePublished - Jul 28 2009

Keywords

  • H
  • Intracellular pH
  • Liposomes
  • Oocyte
  • Xenopus laevis

ASJC Scopus subject areas

  • General

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