The closed structure of the MscS mechanosensitive channel. Cross-linking of single cysteine mutants

Samantha Miller, Michelle Diane Edwards, C. Ozdemir, Ian Rylance Booth

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Mechanosensitive channels must make a large conformational change during the transition from the closed to the open state. The crystal structure of the open form of the Escherichia coli MscS channel was recently solved and depicts a homoheptamer (1). In this study, cross-linking of site-specific cysteine substitutions demonstrates that residues up to 10 - 33 Angstrom apart in the crystal structure readily form disulfide bridges in the closed form and can also be cross-linked by a 10-Angstrom linker. Crosslinking between adjacent subunits stabilizes the heptameric form of the channel providing biochemical evidence to support the crystal structure. The data are consistent with the published model ( 1) in that the membrane domain is highly flexible and that the closed to open transition may involve a significant displacement of transmembrane helices 1 and 2, possibly by as much as 30 Angstrom. The data are also consistent with significant flexibility of the cytoplasmic domain.

Original languageEnglish
Pages (from-to)32246-32250
Number of pages4
JournalThe Journal of Biological Chemistry
Volume278
Issue number34
DOIs
Publication statusPublished - Aug 2003

Keywords

  • ESCHERICHIA-COLI MSCS
  • GATING MECHANISM
  • ION-CHANNEL
  • K+ CHANNEL
  • STOICHIOMETRY
  • RECEPTOR
  • DYNAMICS
  • DOMAIN

Cite this

The closed structure of the MscS mechanosensitive channel. Cross-linking of single cysteine mutants. / Miller, Samantha; Edwards, Michelle Diane; Ozdemir, C.; Booth, Ian Rylance.

In: The Journal of Biological Chemistry, Vol. 278, No. 34, 08.2003, p. 32246-32250.

Research output: Contribution to journalArticle

Miller, Samantha ; Edwards, Michelle Diane ; Ozdemir, C. ; Booth, Ian Rylance. / The closed structure of the MscS mechanosensitive channel. Cross-linking of single cysteine mutants. In: The Journal of Biological Chemistry. 2003 ; Vol. 278, No. 34. pp. 32246-32250.
@article{9cabc91d9a0845078db0985738e7b707,
title = "The closed structure of the MscS mechanosensitive channel. Cross-linking of single cysteine mutants",
abstract = "Mechanosensitive channels must make a large conformational change during the transition from the closed to the open state. The crystal structure of the open form of the Escherichia coli MscS channel was recently solved and depicts a homoheptamer (1). In this study, cross-linking of site-specific cysteine substitutions demonstrates that residues up to 10 - 33 Angstrom apart in the crystal structure readily form disulfide bridges in the closed form and can also be cross-linked by a 10-Angstrom linker. Crosslinking between adjacent subunits stabilizes the heptameric form of the channel providing biochemical evidence to support the crystal structure. The data are consistent with the published model ( 1) in that the membrane domain is highly flexible and that the closed to open transition may involve a significant displacement of transmembrane helices 1 and 2, possibly by as much as 30 Angstrom. The data are also consistent with significant flexibility of the cytoplasmic domain.",
keywords = "ESCHERICHIA-COLI MSCS, GATING MECHANISM, ION-CHANNEL, K+ CHANNEL, STOICHIOMETRY, RECEPTOR, DYNAMICS, DOMAIN",
author = "Samantha Miller and Edwards, {Michelle Diane} and C. Ozdemir and Booth, {Ian Rylance}",
year = "2003",
month = "8",
doi = "10.1074/jbc.M303188200",
language = "English",
volume = "278",
pages = "32246--32250",
journal = "The Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC",
number = "34",

}

TY - JOUR

T1 - The closed structure of the MscS mechanosensitive channel. Cross-linking of single cysteine mutants

AU - Miller, Samantha

AU - Edwards, Michelle Diane

AU - Ozdemir, C.

AU - Booth, Ian Rylance

PY - 2003/8

Y1 - 2003/8

N2 - Mechanosensitive channels must make a large conformational change during the transition from the closed to the open state. The crystal structure of the open form of the Escherichia coli MscS channel was recently solved and depicts a homoheptamer (1). In this study, cross-linking of site-specific cysteine substitutions demonstrates that residues up to 10 - 33 Angstrom apart in the crystal structure readily form disulfide bridges in the closed form and can also be cross-linked by a 10-Angstrom linker. Crosslinking between adjacent subunits stabilizes the heptameric form of the channel providing biochemical evidence to support the crystal structure. The data are consistent with the published model ( 1) in that the membrane domain is highly flexible and that the closed to open transition may involve a significant displacement of transmembrane helices 1 and 2, possibly by as much as 30 Angstrom. The data are also consistent with significant flexibility of the cytoplasmic domain.

AB - Mechanosensitive channels must make a large conformational change during the transition from the closed to the open state. The crystal structure of the open form of the Escherichia coli MscS channel was recently solved and depicts a homoheptamer (1). In this study, cross-linking of site-specific cysteine substitutions demonstrates that residues up to 10 - 33 Angstrom apart in the crystal structure readily form disulfide bridges in the closed form and can also be cross-linked by a 10-Angstrom linker. Crosslinking between adjacent subunits stabilizes the heptameric form of the channel providing biochemical evidence to support the crystal structure. The data are consistent with the published model ( 1) in that the membrane domain is highly flexible and that the closed to open transition may involve a significant displacement of transmembrane helices 1 and 2, possibly by as much as 30 Angstrom. The data are also consistent with significant flexibility of the cytoplasmic domain.

KW - ESCHERICHIA-COLI MSCS

KW - GATING MECHANISM

KW - ION-CHANNEL

KW - K+ CHANNEL

KW - STOICHIOMETRY

KW - RECEPTOR

KW - DYNAMICS

KW - DOMAIN

U2 - 10.1074/jbc.M303188200

DO - 10.1074/jbc.M303188200

M3 - Article

VL - 278

SP - 32246

EP - 32250

JO - The Journal of Biological Chemistry

JF - The Journal of Biological Chemistry

SN - 0021-9258

IS - 34

ER -