The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel

Ken Ichi Hashimoto; Koji Nakamura; Teruo Kuroda; Isamu Yabe; Tsuyoshi Nakamatsu; Hisashi Kawasaki

doi:10.1271/bbb.100636

The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel

Ken Ichi Hashimoto, Koji Nakamura, Teruo Kuroda, Isamu Yabe, Tsuyoshi Nakamatsu, Hisashi Kawasaki

Faculty of Pharmaceutical Sciences

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

32 Citations (Scopus)

Abstract

The function of the NCgl1221-encoded protein of Corynebacterium glutamicum was analyzed using Bacillus subtilis as host because a method for preparing the giant provacuole required for electrophysiological studies has been established. Expression of NCgl1221 in a strain deficient in mscL and ykuT, both of which encode mechanosensitive channels, resulted in an 8.9-fold higher cell survival rate upon osmotic downshock than the control. Electrophysiological investigation showed that the giant provacuole prepared from this strain, expressing NCgl1221, exhibited significantly higher pressuredependent conductance than the control. These findings show that the NCgl1221-encoded protein functions as a mechanosensitive channel.

Original language	English
Pages (from-to)	2546-2549
Number of pages	4
Journal	Bioscience, Biotechnology and Biochemistry
Volume	74
Issue number	12
DOIs	https://doi.org/10.1271/bbb.100636
Publication status	Published - 2010

Keywords

Corynebacterium glutamicum
Electrophysiological study
Glutamate overproduction
Mechanosensitive channel
Patch-clamp method

ASJC Scopus subject areas

Biotechnology
Analytical Chemistry
Biochemistry
Applied Microbiology and Biotechnology
Molecular Biology
Organic Chemistry

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10.1271/bbb.100636

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title = "The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel",

abstract = "The function of the NCgl1221-encoded protein of Corynebacterium glutamicum was analyzed using Bacillus subtilis as host because a method for preparing the giant provacuole required for electrophysiological studies has been established. Expression of NCgl1221 in a strain deficient in mscL and ykuT, both of which encode mechanosensitive channels, resulted in an 8.9-fold higher cell survival rate upon osmotic downshock than the control. Electrophysiological investigation showed that the giant provacuole prepared from this strain, expressing NCgl1221, exhibited significantly higher pressuredependent conductance than the control. These findings show that the NCgl1221-encoded protein functions as a mechanosensitive channel.",

keywords = "Corynebacterium glutamicum, Electrophysiological study, Glutamate overproduction, Mechanosensitive channel, Patch-clamp method",

author = "Hashimoto, {Ken Ichi} and Koji Nakamura and Teruo Kuroda and Isamu Yabe and Tsuyoshi Nakamatsu and Hisashi Kawasaki",

note = "Funding Information: We are grateful to Dr. A. Nakamura, Dr. F. Kawamura, and Dr. S. Kosono for providing pJM116-Pxyl, the erythromycin-resistant gene, and the neomycin-resistant gene respectively. We would also like to thank Dr. G. Akanuma for technical support. This work was supported financially in part by a grant from the Strategic Research Foundation Grant-Aided Project for Private Universities of the Ministry of Education, Culture, Sport, Science, and Technology of Japan (MEXT), 2008–2012 (S0801023).",

year = "2010",

doi = "10.1271/bbb.100636",

language = "English",

volume = "74",

pages = "2546--2549",

journal = "Bioscience, Biotechnology and Biochemistry",

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publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",

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T1 - The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel

AU - Hashimoto, Ken Ichi

AU - Nakamura, Koji

AU - Kuroda, Teruo

AU - Yabe, Isamu

AU - Nakamatsu, Tsuyoshi

AU - Kawasaki, Hisashi

N1 - Funding Information: We are grateful to Dr. A. Nakamura, Dr. F. Kawamura, and Dr. S. Kosono for providing pJM116-Pxyl, the erythromycin-resistant gene, and the neomycin-resistant gene respectively. We would also like to thank Dr. G. Akanuma for technical support. This work was supported financially in part by a grant from the Strategic Research Foundation Grant-Aided Project for Private Universities of the Ministry of Education, Culture, Sport, Science, and Technology of Japan (MEXT), 2008–2012 (S0801023).

PY - 2010

Y1 - 2010

N2 - The function of the NCgl1221-encoded protein of Corynebacterium glutamicum was analyzed using Bacillus subtilis as host because a method for preparing the giant provacuole required for electrophysiological studies has been established. Expression of NCgl1221 in a strain deficient in mscL and ykuT, both of which encode mechanosensitive channels, resulted in an 8.9-fold higher cell survival rate upon osmotic downshock than the control. Electrophysiological investigation showed that the giant provacuole prepared from this strain, expressing NCgl1221, exhibited significantly higher pressuredependent conductance than the control. These findings show that the NCgl1221-encoded protein functions as a mechanosensitive channel.

AB - The function of the NCgl1221-encoded protein of Corynebacterium glutamicum was analyzed using Bacillus subtilis as host because a method for preparing the giant provacuole required for electrophysiological studies has been established. Expression of NCgl1221 in a strain deficient in mscL and ykuT, both of which encode mechanosensitive channels, resulted in an 8.9-fold higher cell survival rate upon osmotic downshock than the control. Electrophysiological investigation showed that the giant provacuole prepared from this strain, expressing NCgl1221, exhibited significantly higher pressuredependent conductance than the control. These findings show that the NCgl1221-encoded protein functions as a mechanosensitive channel.

KW - Corynebacterium glutamicum

KW - Electrophysiological study

KW - Glutamate overproduction

KW - Mechanosensitive channel

KW - Patch-clamp method

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The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel

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