Temperature-sensitive mutants of MscL mechanosensitive channel

Naoto Owada, Megumi Yoshida, Kohei Morita, Kenjiro Yoshimura

Research output: Contribution to journalArticle

Abstract

MscL is a mechanosensitive channel that undergoes a global conformational change upon application of membrane stretching. To elucidate how the structural stability and flexibility occur, we isolated temperature-sensitive (Ts) mutants of Escherichia coli MscL that allowed cell growth at 32°C but not at 42°C. Two Ts mutants, L86P and D127V, were identified. The L86P mutation occurred in the second transmembrane helix, TM2. Substitution of residues neighbouring L86 with proline also led to a Ts mutation, but the substitution of L86 with other amino acids did not result in a Ts phenotype, indicating that the Ts phenotype was due to a structural change of TM2 helix by the introduction of a proline residue. The D127V mutation was localized in the electrostatic belt of the bundle of cytoplasmic helices, indicating that stability of the pentameric bundle of the cytoplasmic helix affects MscL structure. Together, this study described a novel class of MscL mutations that were correlated with the thermodynamic stability of the MscL structure.

Original languageEnglish
Pages (from-to)281-288
Number of pages8
JournalJournal of biochemistry
Volume166
Issue number3
DOIs
Publication statusPublished - 2019 Sep 1

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Proline
Substitution reactions
Temperature
Cell growth
Escherichia coli
Stretching
Electrostatics
Thermodynamic stability
Membranes
Amino Acids

Keywords

  • bacteria
  • gain-of-function
  • mechanosensitive channel
  • temperature-sensitive mutants
  • αhelix

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Temperature-sensitive mutants of MscL mechanosensitive channel. / Owada, Naoto; Yoshida, Megumi; Morita, Kohei; Yoshimura, Kenjiro.

In: Journal of biochemistry, Vol. 166, No. 3, 01.09.2019, p. 281-288.

Research output: Contribution to journalArticle

Owada, Naoto ; Yoshida, Megumi ; Morita, Kohei ; Yoshimura, Kenjiro. / Temperature-sensitive mutants of MscL mechanosensitive channel. In: Journal of biochemistry. 2019 ; Vol. 166, No. 3. pp. 281-288.
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