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The Journal of Membrane Biology

, Volume 247, Issue 9–10, pp 843–851 | Cite as

Application of Amphipols for Structure–Functional Analysis of TRP Channels

  • Kevin W. Huynh
  • Matthew R. Cohen
  • Vera Y. Moiseenkova-BellEmail author
Article

Abstract

Amphipathic polymers (amphipols), such as A8-35 and SApol, are a new tool for stabilizing integral membrane proteins in detergent-free conditions for structural and functional studies. Transient receptor potential (TRP) ion channels function as tetrameric protein complexes in a diverse range of cellular processes including sensory transduction. Mammalian TRP channels share ~20 % sequence similarity and are categorized into six subfamilies: TRPC (canonical), TRPV (vanilloid), TRPA (ankyrin), TRPM (melastatin), TRPP (polycystin), and TRPML (mucolipin). Due to the inherent difficulties in purifying eukaryotic membrane proteins, structural studies of TRP channels have been limited. Recently, A8-35 was essential in resolving the molecular architecture of the nociceptor TRPA1 and led to the determination of a high-resolution structure of the thermosensitive TRPV1 channel by cryo-EM. Newly developed maltose-neopentyl glycol (MNG) detergents have also proven to be useful in stabilizing TRP channels for structural analysis. In this review, we will discuss the impacts of amphipols and MNG detergents on structural studies of TRP channels by cryo-EM. We will compare how A8-35 and MNG detergents interact with the hydrophobic transmembrane domains of TRP channels. In addition, we will discuss what these cryo-EM studies reveal on the importance of screening different types of surfactants toward determining high-resolution structures of TRP channels.

Keywords

Cryo-EM Amphipols Detergent TRPA1 TRPV1 TRPV2 

Abbreviations

A8-35

A poly(sodium acrylate)-based amphipol compromising 35 % of free carboxylates, 25 % of octyl chains, 40 % of isopropyl groups

SApol

Sulfonated amphipol

DM

Decyl-β-D-maltoside

DDM

Dodecyl-β-D-maltoside

MNG

Maltose-neopentyl glycol

2D

2-Dimensional

TM

Transmembrane

TRPA

Transient receptor potential ankyrin

TRPV

Transient receptor potential vanilloid

EM

Electron microscopy

SEC

Size-exclusion chromatography

Notes

Acknowledgments

We would like to thank Teresa Cvetkov for her contribution to this project. We are also very grateful to Jean-Luc Popot for providing us with amphipols and helpful discussions. This work was supported by the American Heart Association (NCRP Scientist Development Grant 11SDG5280029), the American Lung Association Biomedical Research Grant (RG-166985-N), and the National Institute of Health Grant (NIGMS 1R01GM103899-01A1).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Kevin W. Huynh
    • 1
  • Matthew R. Cohen
    • 2
  • Vera Y. Moiseenkova-Bell
    • 1
    • 2
    Email author
  1. 1.Department of Pharmacology, School of MedicineCase Western Reserve UniversityClevelandUSA
  2. 2.Department of Physiology and Biophysics, School of MedicineCase Western Reserve UniversityClevelandUSA

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