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...

Full description

Saved in:
Bibliographic Details
Published in:FEBS letters 2009-01, Vol.583 (2), p.407-412
Main Authors: Battle, Andrew R., Petrov, Evgeny, Pal, Prithwish, Martinac, Boris
Format: Article
Language:English
Subjects:
Bacteria
Cell Membrane - physiology
Confocal microscopy
Escherichia coli Proteins - chemistry
Escherichia coli Proteins - genetics
Escherichia coli Proteins - physiology
Fluorescence
Ion Channels - chemistry
Ion Channels - genetics
Ion Channels - physiology
Lipid
Mechanotransduction, Cellular
Methods
Microscopy, Confocal
Patch clamp
Phosphatidylcholines - chemistry
Sucrose - chemistry
Sugar
Unilamellar Liposomes - chemistry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 6 h) 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.
ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2008.12.033