Engineering covalent oligomers of the mechanosensitive channel of large conductance from Escherichia coli with native conductance and gating characteristics

JHA Folgering, JC Wolters, B Poolman*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)
189 Downloads (Pure)

Abstract

To obtain a gene construct for making single substitutions per channel and to determine the quaternary structure of the mechanosensitive channel MscL from Escherichia coli, covalent oligomers (monomer to hexamer) were engineered by gene fusion; up to six copies of the mscL gene were fused in tandem. All the multimeric tandem constructs yielded functional channels with wild-type conductance and dwell times. Importantly, only the covalent pentamer opened at the same relative pressure (compared to the pressure required to open MscS) as the wild-type MscL channel. The in vivo data strongly suggest that pentameric MscL represents the functional state of the channel.

Original languageEnglish
Pages (from-to)2947-2954
Number of pages8
JournalProtein Science
Volume14
Issue number12
DOIs
Publication statusPublished - Dec-2005

Keywords

  • MscL
  • oligomeric structure
  • covalently linked oligomer
  • structure/function studies
  • membrane proteins
  • ION-CHANNEL
  • FUNCTIONAL RECONSTITUTION
  • SINGLE RESIDUE
  • MSCL
  • PROTEIN
  • STOICHIOMETRY
  • PURIFICATION

Cite this