title: The gating mechanism of the large mechanosensitive channel MscL.
abstract: The mechanosensitive channel of large conductance, MscL, is a ubiquitous membrane-embedded valve involved in turgor regulation in bacteria. The crystal structure of MscL from Mycobacterium tuberculosis provides a starting point for analysing molecular mechanisms of tension-dependent channel gating. Here we develop structural models in which a cytoplasmic gate is formed by a bundle of five amino-terminal helices (S1), previously unresolved in the crystal structure. When membrane tension is applied, the transmembrane barrel expands and pulls the gate apart through the S1-M1 linker. We tested these models by substituting cysteines for residues predicted to be near each other only in either the closed or open conformation. Our results demonstrate that S1 segments form the bundle when the channel is closed, and crosslinking between S1 segments prevents opening. S1 segments interact with M2 when the channel is open, and crosslinking of S1 to M2 impedes channel closing. Gating is affected by the length of the S1-M1 linker in a manner consistent with the model, revealing critical spatial relationships between the domains that transmit force from the lipid bilayer to the channel gate.
authors.affiliation: Department of Biology, University of Maryland, College Park 20742, USA. ss311@umail.umd.edu
authors.forename: S
authors.initials: S
authors.lastname: Sukharev
authors.forename: M
authors.initials: M
authors.lastname: Betanzos
authors.forename: C S
authors.initials: CS
authors.lastname: Chiang
authors.forename: H R
authors.initials: HR
authors.lastname: Guy
mesh_terms: D001696:Biomechanical Phenomena; D003545:Cysteine / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D029968:Escherichia coli Proteins; D015640:Ion Channel Gating; D007473:Ion Channels / Q000378:metabolism*; D008954:Models, Biological; D008958:Models, Molecular; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D004220:Disulfides; D029968:Escherichia coli Proteins; D007473:Ion Channels; C086596:MscL protein, E coli; D003545:Cysteine