title: Lipid-protein interaction of the MscS mechanosensitive channel examined by scanning mutagenesis.
abstract: The mechanosensitive channel of small conductance (MscS) is a bacterial mechanosensitive channel that opens in response to rapid hypoosmotic stress. Since MscS can be opened solely by membrane stretch without help from any accessory protein, the lipid-protein interface must play a crucial role in sensing membrane tension. In this study, the hydrophobic residues in the lipid-protein interface were substituted one by one with a hydrophilic amino acid, asparagine, to modify the interaction between the protein and the lipid. Function of the mutant MscSs was examined by patch-clamp and hypoosmotic shock experiments. An increase in the gating threshold and a decrease in the viability on hypoosmotic shock were observed when the hydrophobic residues near either end of the first or the second transmembrane helix (TM1 or TM2) were replaced with asparagine. This observation indicates that the lipid-protein interaction at the ends of both helices (TM1 and TM2) is essential to MscS function.
authors.affiliation: ICORP/SORST, Cell Mechanosensing, Japan Science and Technology Agency, Nagoya, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nomura
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sokabe
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Yoshimura
mesh_terms: D019943:Amino Acid Substitution; D001216:Asparagine / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000502:physiology*; D057927:Hydrophobic and Hydrophilic Interactions; D007473:Ion Channels / Q000235:genetics / Q000502:physiology*; D040542:Mechanotransduction, Cellular; D008563:Membrane Lipids / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D009997:Osmotic Pressure; D018408:Patch-Clamp Techniques; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013104:Spheroplasts / Q000235:genetics / Q000502:physiology
chemical_list: D029968:Escherichia coli Proteins; D007473:Ion Channels; D008563:Membrane Lipids; C469542:MscS protein, E coli; D001216:Asparagine