Structural and functional characterization of an otopetrin family proton channel

Qingfeng Chen, Weizhong Zeng, Ji She, Xiaochen Bai, Youxing Jiang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The otopetrin (OTOP) proteins were recently characterized as proton channels. Here we present the cryo-EM structure of OTOP3 from Xenopus tropicalis (XtOTOP3) along with functional characterization of the channel. XtOTOP3 forms a homodimer with each subunit containing 12 transmembrane helices that can be divided into two structurally homologous halves; each half assembles as an α-helical barrel that could potentially serve as a proton conduction pore. Both pores open from the extracellular half before becoming occluded at a central constriction point consisting of three highly conserved residues - Gln232/585-Asp262/Asn623-Tyr322/666 (the constriction triads). Mutagenesis shows that the constriction triad from the second pore is less amenable to perturbation than that of the first pore, suggesting an unequal contribution between the two pores to proton transport. We also identified several key residues at the interface between the two pores that are functionally important, particularly Asp509, which confers intracellular pH-dependent desensitization to OTOP channels.

Original languageEnglish (US)
JournaleLife
Volume8
DOIs
StatePublished - Apr 11 2019

Fingerprint

Constriction
Protons
Mutagenesis
Xenopus
Proteins

Keywords

  • cryo-EM
  • membrane channels
  • molecular biophysics
  • otopetrin
  • proton
  • structural biology
  • xenopus

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Structural and functional characterization of an otopetrin family proton channel. / Chen, Qingfeng; Zeng, Weizhong; She, Ji; Bai, Xiaochen; Jiang, Youxing.

In: eLife, Vol. 8, 11.04.2019.

Research output: Contribution to journalArticle

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N2 - The otopetrin (OTOP) proteins were recently characterized as proton channels. Here we present the cryo-EM structure of OTOP3 from Xenopus tropicalis (XtOTOP3) along with functional characterization of the channel. XtOTOP3 forms a homodimer with each subunit containing 12 transmembrane helices that can be divided into two structurally homologous halves; each half assembles as an α-helical barrel that could potentially serve as a proton conduction pore. Both pores open from the extracellular half before becoming occluded at a central constriction point consisting of three highly conserved residues - Gln232/585-Asp262/Asn623-Tyr322/666 (the constriction triads). Mutagenesis shows that the constriction triad from the second pore is less amenable to perturbation than that of the first pore, suggesting an unequal contribution between the two pores to proton transport. We also identified several key residues at the interface between the two pores that are functionally important, particularly Asp509, which confers intracellular pH-dependent desensitization to OTOP channels.

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