Surface changes of the mechanosensitive channel MscS upon its activation, inactivation, and closing.
MscS is a bacterial mechanosensitive channel that shows voltage dependence. The crystal structure of MscS revealed that the channel is a homoheptamer with a large chamber on the intracellular site. Our previous experiments indicated that the cytoplasmic chamber of the channel is not a rigid structure and changes its conformation upon the channel activation. In this study, we have applied various sized cosolvents that are excluded from protein surfaces. It is well known that such cosolvents induce compaction of proteins and prevent thermal fluctuations. It is also known that they shift channel equilibrium to the state of lower volume. We have found that large cosolvents that cannot enter the channel interior accelerate channel inactivation when applied from the cytoplasmic side, but they slow down inactivation when applied from the extracellular side. We have also found that small cosolvents that can enter the channel cytoplasmic chamber prevent the channel from opening, unlike the large ones. These data support our idea that the channel cytoplasmic chamber shrinks upon inactivation but also give new clues about conformational changes of the channel upon transitions between its functional states.
Grajkowski
Wojciech
W
Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland.
Kubalski
Andrzej
A
Koprowski
Piotr
P
D003593:Cytoplasm / Q000378:metabolism; D003911:Dextrans / Q000737:chemistry; D004594:Electrophysiology; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry*; D005362:Ficoll / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D015640:Ion Channel Gating; D007473:Ion Channels / Q000737:chemistry*; D040542:Mechanotransduction, Cellular; D008958:Models, Molecular; D011092:Polyethylene Glycols / Q000737:chemistry; D011312:Pressure; D011485:Protein Binding; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D011506:Proteins / Q000737:chemistry; D011523:Protoplasts / Q000378:metabolism; D013499:Surface Properties; D013997:Time Factors
D003911:Dextrans; D029968:Escherichia coli Proteins; D007473:Ion Channels; C469542:MscS protein, E coli; D011506:Proteins; D005362:Ficoll; D011092:Polyethylene Glycols; D005947:Glucose