Rapid and improved reconstitution of bacterial mechanosensitive ion channel proteins MscS and MscL into liposomes using a modified sucrose method.
The bacterial mechanosensitive (MS) channels of small (MscS) and large (MscL) conductance have functionally been reconstituted into giant unilamellar liposomes (GUVs) using an improved reconstitution method in the presence of sucrose. This method gives significant time savings (preparation times as little as 6h) compared to the classical method of protein reconstitution which uses a dehydration/rehydration (D/R) procedure (minimum 2 days preparation time). Moreover, it represents the first highly reproducible method for functional reconstitution of MscS as well as MscS/MscL co-reconstitution. This novel procedure has the potential to be used for studies of other ion channels by liposome reconstitution.
Battle
Andrew R
AR
Department of Physiology and Pharmacology, School of Biomedical Sciences, The University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia.
Petrov
Evgeny
E
Pal
Prithwish
P
Martinac
Boris
B
D002462:Cell Membrane / Q000502:physiology; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D007473:Ion Channels / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D040542:Mechanotransduction, Cellular; D008722:Methods; D018613:Microscopy, Confocal; D010713:Phosphatidylcholines / Q000737:chemistry; D013395:Sucrose / Q000737:chemistry*; D053835:Unilamellar Liposomes / Q000737:chemistry*
D029968:Escherichia coli Proteins; D007473:Ion Channels; C086596:MscL protein, E coli; C469542:MscS protein, E coli; D010713:Phosphatidylcholines; D053835:Unilamellar Liposomes; D013395:Sucrose; C024990:asolectin