An in vivo assay identifies changes in residue accessibility on mechanosensitive channel gating

Proceedings of the National Academy of Sciences of the United States of America
Jessica L BartlettPaul Blount

Abstract

MscL is a mechanosensitive channel of large conductance that functions as an "emergency release valve," allowing bacteria to survive acute hypoosmotic stress. Although Escherichia coli MscL is the best-studied mechanosensitive channel, structural rearrangements occurring during gating remain disputed. Introduction of a charged residue into the pore of MscL was shown to result in a reduced-viability phenotype. Here, we probe for residues in the transmembrane domains that are exposed to the aqueous environment in the presence and absence of hypoosmotic shock by reacting a charged sulfhydryl reagent with substituted cysteines. Subsequent analysis of cell viability allows for an assessment of residues exposed in the closed and opening states in vivo. The results suggest that the crystal structure of MscL derived from the Mycobacterium tuberculosis orthologue may reflect a nearly closed rather than fully closed state and support a clockwise rotation of the pore-forming first transmembrane domain on gating.

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Citations

May 9, 2009·European Biophysics Journal : EBJ·Daniel Balleza, Froylan Gómez-Lagunas
Dec 31, 2011·ACS Nano·Li-Min YangPaul Blount
Feb 11, 2005·Nature Structural & Molecular Biology·Boris Martinac
Jan 25, 2005·Nature Structural & Molecular Biology·Michelle D EdwardsIan R Booth
Apr 23, 2009·The Journal of Biological Chemistry·Andriy Anishkin, Sergei Sukharev
Feb 3, 2005·The Journal of General Physiology·Andriy AnishkinSergei Sukharev
Sep 30, 2010·The Journal of General Physiology·Ben CorryBoris Martinac
Mar 6, 2009·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Yuezhou LiPaul Blount
Oct 12, 2010·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Li-Min Yang, Paul Blount
Nov 12, 2010·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Irene IsclaPaul Blount
Mar 13, 2012·Annual Review of Biophysics·James H Naismith, Ian R Booth
Sep 11, 2014·Nature Communications·Irene IsclaPaul Blount
Aug 12, 2015·Proceedings of the National Academy of Sciences of the United States of America·Jie LiZhenfeng Liu
Jan 24, 2008·Biophysical Journal·Jonggu Jeon, Gregory A Voth
Jul 12, 2005·Current Opinion in Neurobiology·Andriy Anishkin, Ching Kung
Nov 23, 2006·Biophysical Journal·Irene IsclaPaul Blount
Nov 19, 2014·Proceedings of the National Academy of Sciences of the United States of America·Albert KonijnenbergArmağan Koçer
May 17, 2013·Biosensors·Irene IsclaPaul Blount
Oct 26, 2018·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Robin WrayPaul Blount
May 31, 2019·Journal of Biochemistry·Naoto OwadaKenjiro Yoshimura
Jan 17, 2020·Microbiology and Molecular Biology Reviews : MMBR·Paul Blount, Irene Iscla
Oct 18, 2014·Physical Review Letters·Douwe Jan Bonthuis, Ramin Golestanian
Dec 23, 2004·The Journal of Biological Chemistry·Joost H A FolgeringBert Poolman
Aug 1, 2009·EcoSal Plus·Karlheinz AltendorfJanet M Wood

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