X-ray crystallographic structure of thermophilic rhodopsin: Implications For High Thermal Stability And Optogenetil Function

Takashi Tsukamoto, Kenji Mizutani, Taisuke Hasegawa, Megumi Takahashi, Naoya Honda, Naoki Hashimoto, Kazumi Shimono, Keitaro Yamashita, Masaki Yamamoto, Seiji Miyauchi, Shin Takagi, Shigehiko Hayashi, Takeshi Murata, Yuki Sudo

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Thermophilic rhodopsin (TR) is a photoreceptor protein with an extremely high thermal stability and the first characterized light-driven electrogenic proton pump derived from the extreme thermophile Thermus thermophilus JL-18. In this study, we confirmed its high thermal stability compared with other microbial rhodopsins and also report the potential availability of TR for optogenetics as a light-induced neural silencer. The x-ray crystal structure of TR revealed that its overall structure is quite similar to that of xanthorhodopsin, including the presence of a putative binding site for a carotenoid antenna; but several distinct structural characteristics of TR, including a decreased surface charge and a larger number of hydrophobic residues and aromatic-aromatic interactions, were also clarified. Based on the crystal structure, the structural changes of TR upon thermal stimulation were investigated by molecular dynamics simulations. The simulations revealed the presence of a thermally induced structural substate in which an increase of hydrophobic interactions in the extracellular domain, the movement of extracellular domains, the formation of a hydrogen bond, and the tilting of transmembrane helices were observed. From the computational and mutational analysis, we propose that an extracellular LPGG motif between helices F and G plays an important role in the thermal stability, acting as a "thermal sensor." These findings will be valuable for understanding retinal proteins with regard to high protein stability and high optogenetic performance.

Original languageEnglish
Pages (from-to)12223-12232
Number of pages10
JournalJournal of Biological Chemistry
Volume291
Issue number23
DOIs
Publication statusPublished - Jun 3 2016

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Rhodopsin
Thermodynamic stability
Hot Temperature
X-Rays
X rays
Optogenetics
Microbial Rhodopsins
Crystal structure
Thermus thermophilus
Light
Proton Pumps
Proteins
Protein Stability
Molecular Dynamics Simulation
Carotenoids
Surface charge
Hydrophobic and Hydrophilic Interactions
Molecular dynamics
Hydrogen
Hydrogen bonds

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

X-ray crystallographic structure of thermophilic rhodopsin : Implications For High Thermal Stability And Optogenetil Function. / Tsukamoto, Takashi; Mizutani, Kenji; Hasegawa, Taisuke; Takahashi, Megumi; Honda, Naoya; Hashimoto, Naoki; Shimono, Kazumi; Yamashita, Keitaro; Yamamoto, Masaki; Miyauchi, Seiji; Takagi, Shin; Hayashi, Shigehiko; Murata, Takeshi; Sudo, Yuki.

In: Journal of Biological Chemistry, Vol. 291, No. 23, 03.06.2016, p. 12223-12232.

Research output: Contribution to journalArticle

Tsukamoto, T, Mizutani, K, Hasegawa, T, Takahashi, M, Honda, N, Hashimoto, N, Shimono, K, Yamashita, K, Yamamoto, M, Miyauchi, S, Takagi, S, Hayashi, S, Murata, T & Sudo, Y 2016, 'X-ray crystallographic structure of thermophilic rhodopsin: Implications For High Thermal Stability And Optogenetil Function', Journal of Biological Chemistry, vol. 291, no. 23, pp. 12223-12232. https://doi.org/10.1074/jbc.M116.719815
Tsukamoto, Takashi ; Mizutani, Kenji ; Hasegawa, Taisuke ; Takahashi, Megumi ; Honda, Naoya ; Hashimoto, Naoki ; Shimono, Kazumi ; Yamashita, Keitaro ; Yamamoto, Masaki ; Miyauchi, Seiji ; Takagi, Shin ; Hayashi, Shigehiko ; Murata, Takeshi ; Sudo, Yuki. / X-ray crystallographic structure of thermophilic rhodopsin : Implications For High Thermal Stability And Optogenetil Function. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 23. pp. 12223-12232.
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