Engineered Functional Recovery of Microbial Rhodopsin Without Retinal-Binding Lysine

Yumeka Yamauchi; Masae Konno; Daichi Yamada; Kei Yura; Keiichi Inoue; Oded Beja; Hideki Kandori

doi:10.1111/php.13114

Engineered Functional Recovery of Microbial Rhodopsin Without Retinal-Binding Lysine

Yumeka Yamauchi, Masae Konno, Daichi Yamada, Kei Yura, Keiichi Inoue, Oded Beja, Hideki Kandori

Technion - Israel Institute of Technology

Research output: Contribution to journal › Article › peer-review

Abstract

Definition of rhodopsin is the retinal-binding membrane protein with the Schiff base linkage at a lysine on the 7^th transmembrane helix. However, ~ 600 microbial rhodopsins lack retinal-binding lysine at the corresponding position (Rh-noK) among ~ 5500 known microbial rhodopsins, suggesting that Rh-noK has each functional role without chromophore. Here, we report successful functional recovery of Rh-noK. Two Rh-noKs from bacteria were heterologously expressed in Escherichia coli, which exhibited no color. When retinal-binding lysine was introduced, one of them gained visible color. Additional mutation of the Schiff base counterion further gained proton-pumping activity. Successful engineered functional recovery such as visible color and proton-pump activity suggests that the Rh-noK protein forms a characteristic structure of microbial rhodopsins.

Original language	English
Pages (from-to)	1116-1121
Number of pages	6
Journal	Photochemistry and Photobiology
Volume	95
Issue number	5
DOIs	https://doi.org/10.1111/php.13114
State	Published - 1 Sep 2019

All Science Journal Classification (ASJC) codes

Biochemistry
Physical and Theoretical Chemistry

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10.1111/php.13114

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title = "Engineered Functional Recovery of Microbial Rhodopsin Without Retinal-Binding Lysine",

abstract = "Definition of rhodopsin is the retinal-binding membrane protein with the Schiff base linkage at a lysine on the 7th transmembrane helix. However, \textasciitilde{} 600 microbial rhodopsins lack retinal-binding lysine at the corresponding position (Rh-noK) among \textasciitilde{} 5500 known microbial rhodopsins, suggesting that Rh-noK has each functional role without chromophore. Here, we report successful functional recovery of Rh-noK. Two Rh-noKs from bacteria were heterologously expressed in Escherichia coli, which exhibited no color. When retinal-binding lysine was introduced, one of them gained visible color. Additional mutation of the Schiff base counterion further gained proton-pumping activity. Successful engineered functional recovery such as visible color and proton-pump activity suggests that the Rh-noK protein forms a characteristic structure of microbial rhodopsins.",

author = "Yumeka Yamauchi and Masae Konno and Daichi Yamada and Kei Yura and Keiichi Inoue and Oded Beja and Hideki Kandori",

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AU - Yamauchi, Yumeka

AU - Konno, Masae

AU - Yamada, Daichi

AU - Yura, Kei

AU - Inoue, Keiichi

AU - Beja, Oded

AU - Kandori, Hideki

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N2 - Definition of rhodopsin is the retinal-binding membrane protein with the Schiff base linkage at a lysine on the 7th transmembrane helix. However, ~ 600 microbial rhodopsins lack retinal-binding lysine at the corresponding position (Rh-noK) among ~ 5500 known microbial rhodopsins, suggesting that Rh-noK has each functional role without chromophore. Here, we report successful functional recovery of Rh-noK. Two Rh-noKs from bacteria were heterologously expressed in Escherichia coli, which exhibited no color. When retinal-binding lysine was introduced, one of them gained visible color. Additional mutation of the Schiff base counterion further gained proton-pumping activity. Successful engineered functional recovery such as visible color and proton-pump activity suggests that the Rh-noK protein forms a characteristic structure of microbial rhodopsins.

AB - Definition of rhodopsin is the retinal-binding membrane protein with the Schiff base linkage at a lysine on the 7th transmembrane helix. However, ~ 600 microbial rhodopsins lack retinal-binding lysine at the corresponding position (Rh-noK) among ~ 5500 known microbial rhodopsins, suggesting that Rh-noK has each functional role without chromophore. Here, we report successful functional recovery of Rh-noK. Two Rh-noKs from bacteria were heterologously expressed in Escherichia coli, which exhibited no color. When retinal-binding lysine was introduced, one of them gained visible color. Additional mutation of the Schiff base counterion further gained proton-pumping activity. Successful engineered functional recovery such as visible color and proton-pump activity suggests that the Rh-noK protein forms a characteristic structure of microbial rhodopsins.

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Engineered Functional Recovery of Microbial Rhodopsin Without Retinal-Binding Lysine

Abstract

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