title: A large-conductance mechanosensitive channel in E. coli encoded by mscL alone.
abstract: All cellular organisms respond to vibration, touch, gravity or changes in osmolarity, although the molecules on which such mechanosensations depend are unknown. Candidates include certain channels that gate in response to membrane stretch. Patch-clamp experiments with Escherichia coli envelope have revealed a mechanosensitive channel with very large conductance (MscL) and one with a smaller conductance (MscS) which may be important in osmoregulation. Here we have solubilized and fractionated the envelope, reconstituted the MscL activity in vitro, and traced it to a small protein, whose gene, mscL, we then cloned. Insertional disruption of mscL removes the channel activity, whereas re-expression of mscL borne on an expression plasmid restores it. MscL-channel activities were observed in material from a cell-free expression system with mscL as the only template. The mscL nucleotide sequence predicts a unique protein of only 136 amino acids, with a highly hydrophobic core and very different from porins or other known proteins.
authors.affiliation: Laboratory of Molecular Biology, University of Wisconsin-Madison 53706.
authors.forename: S I
authors.initials: SI
authors.lastname: Sukharev
authors.forename: P
authors.initials: P
authors.lastname: Blount
authors.forename: B
authors.initials: B
authors.lastname: Martinac
authors.forename: F R
authors.initials: FR
authors.lastname: Blattner
authors.forename: C
authors.initials: C
authors.lastname: Kung
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004285:Dogs; D004553:Electric Conductivity; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D007473:Ion Channels / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D012995:Solubility; D013104:Spheroplasts
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D007473:Ion Channels; C086596:MscL protein, E coli