Contributions of the different extramembranous domains of the mechanosensitive ion channel MscL to its response to membrane tension.
MscL is a mechanosensitive channel that is gated by tension in the membrane bilayer alone. It is a homo-oligomer of a protein comprising two transmembrane segments connected by an external loop, with the NH(2) and COOH termini located in the cytoplasm. The contributions of the extramembranous domains of the channel to its activity were investigated by specific proteolysis during patch-clamp experiments. Limited proteolysis of the COOH terminus or the NH(2) terminus increased the mechanosensitivity of the channel without changing its conductance. Strikingly, after cleavage of the external loop of each monomer, the channel was still functional, and its mechanosensitivity was increased dramatically, indicating that the loop acts as a spring that resists the opening of the channel and promotes its closure when it is open. These results indicate that the integrity of most of the extramembranous domains is not essential for mechanosensitivity. They suggest that these domains counteract the movement of the transmembrane helices to which they are connected, thus setting the level of sensitivity of the channel to tension.
Ajouz
B
B
Laboratoire des Biomembranes, Unité Mixte de Recherche CNRS 8619, Bâtiment 430, Université Paris-Sud 91405 Orsay Cedex France.
Berrier
C
C
Besnard
M
M
Martinac
B
B
Ghazi
A
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology; D002918:Chymotrypsin / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D007473:Ion Channels / Q000737:chemistry* / Q000502:physiology*; D008051:Lipid Bilayers; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011510:Proteolipids / Q000378:metabolism*; D011523:Protoplasts / Q000502:physiology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism / Q000494:pharmacology
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; C086596:MscL protein, E coli; D011510:Proteolipids; D011994:Recombinant Proteins; C020485:proteoliposomes; D002918:Chymotrypsin; D014357:Trypsin