The conserved carboxy-terminus of the MscS mechanosensitive channel is not essential but increases stability and activity.
The Escherichia coli MscS mechanosensitive channel protein has a distinct domain structure that terminates in a conserved seven-strand beta barrel. This distinctive feature suggested it could be a critical determinant of channel stability and activity. Measurements on a protein deleted for the base of the vestibule and the beta barrel (residues 266-286) suggested that the modified channel had reduced activity. However, induction of the mutant protein resulted in membrane protein accumulation equivalent to wild type and a physiologically functional channel. In patch clamp analysis the activity profile was similar to wild type but reduced numbers of channel were seen per patch, suggesting reduced assembly or stability of the mutant protein. The mutant channel exhibited a subtle change in character - channels did not re-open after full desensitization. Thus the immediate carboxy-terminus (residues 266-286) is not essential for MscS gating but improves stability and activity and is required for recovery of channel activity after desensitization.
School of Medical Sciences, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK.
Ulrike
U
Schumann
Michelle D
MD
Edwards
Chan
C
Li
Ian R
IR
Booth
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D007473:Ion Channels / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011487:Protein Conformation; D017384:Sequence Deletion
D029968:Escherichia coli Proteins; D007473:Ion Channels; C469542:MscS protein, E coli