Rapid and improved reconstitution of bacterial mechanosensitive ion channel proteins MscS and MscL into liposomes using a modified sucrose method
- PMID: 19111548
- DOI: 10.1016/j.febslet.2008.12.033
Rapid and improved reconstitution of bacterial mechanosensitive ion channel proteins MscS and MscL into liposomes using a modified sucrose method
Abstract
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.
Similar articles
-
Mechanosensitive channel of Thermoplasma, the cell wall-less archaea: cloning and molecular characterization.Cell Biochem Biophys. 2001;34(3):321-47. doi: 10.1385/CBB:34:3:321. Cell Biochem Biophys. 2001. PMID: 11898860
-
Gating the bacterial mechanosensitive channels: MscS a new paradigm?Curr Opin Microbiol. 2004 Apr;7(2):163-7. doi: 10.1016/j.mib.2004.02.006. Curr Opin Microbiol. 2004. PMID: 15063854 Review.
-
MscS, the bacterial mechanosensitive channel of small conductance.Int J Biochem Cell Biol. 2008;40(4):581-5. doi: 10.1016/j.biocel.2007.03.013. Epub 2007 Mar 24. Int J Biochem Cell Biol. 2008. PMID: 17466568 Review.
-
An optimized purification and reconstitution method for the MscS channel: strategies for spectroscopical analysis.Biochemistry. 2007 Jun 12;46(23):6766-73. doi: 10.1021/bi700322k. Epub 2007 May 15. Biochemistry. 2007. PMID: 17500538
-
Crystal structure of Escherichia coli MscS, a voltage-modulated and mechanosensitive channel.Science. 2002 Nov 22;298(5598):1582-7. doi: 10.1126/science.1077945. Science. 2002. PMID: 12446901
Cited by
-
Mechanical coupling of the multiple structural elements of the large-conductance mechanosensitive channel during expansion.Proc Natl Acad Sci U S A. 2015 Aug 25;112(34):10726-31. doi: 10.1073/pnas.1503202112. Epub 2015 Aug 10. Proc Natl Acad Sci U S A. 2015. PMID: 26261325 Free PMC article.
-
Formation of Cell-Sized Liposomes Incorporating a β-Barrel-Structured Porin through Rehydration of a Phospholipid-Membrane Protein Dried Film.ACS Omega. 2024 Jan 26;9(5):5911-5918. doi: 10.1021/acsomega.3c09431. eCollection 2024 Feb 6. ACS Omega. 2024. PMID: 38343955 Free PMC article.
-
Detergent-mediated incorporation of transmembrane proteins in giant unilamellar vesicles with controlled physiological contents.Proc Natl Acad Sci U S A. 2013 Apr 30;110(18):7276-81. doi: 10.1073/pnas.1303857110. Epub 2013 Apr 15. Proc Natl Acad Sci U S A. 2013. PMID: 23589883 Free PMC article.
-
Binding of fullerenes and nanotubes to MscL.Sci Rep. 2014 Jul 17;4:5609. doi: 10.1038/srep05609. Sci Rep. 2014. PMID: 25030051 Free PMC article.
-
In Vitro Synthesis and Reconstitution Using Mammalian Cell-Free Lysates Enables the Systematic Study of the Regulation of LINC Complex Assembly.Biochemistry. 2022 Jul 19;61(14):1495-1507. doi: 10.1021/acs.biochem.2c00118. Epub 2022 Jun 23. Biochemistry. 2022. PMID: 35737522 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
