Elucidating a role for the cytoplasmic domain in the Mycobacterium tuberculosis mechanosensitive channel of large conductance

Herrera, Nadia; Maksaev, Grigory; Haswell, Elizabeth S.; Rees, Douglas C.

doi:10.1038/s41598-018-32536-6

Title: Elucidating a role for the cytoplasmic domain in the Mycobacterium tuberculosis mechanosensitive channel of large conductance

Journal Article · Mon Oct 01 00:00:00 EDT 2018 · Scientific Reports

DOI:https://doi.org/10.1038/s41598-018-32536-6· OSTI ID:1624426

^[1];

^[2];

^[1]

California Institute of Technology (CalTech), Pasadena, CA (United States). Howard Hughes Medical Center, Division of Chemistry and Chemical Engineering
Washington Univ., St. Louis, MO (United States). NSF Center for Engineering Mechanobiology, Dept. of Biology

Microbial survival in dynamic environments requires the ability to successfully respond to abrupt changes in osmolarity. The mechanosensitive channel of large conductance (MscL) is a ubiquitous channel that facilitates the survival of bacteria and archaea under severe osmotic down-shock conditions by relieving excess turgor pressure in response to increased membrane tension. A prominent structural feature of MscL, the cytoplasmic C-terminal domain, has been suggested to influence channel assembly and function. In this report, we describe the X-ray crystal structure and electrophysiological properties of a C-terminal domain truncation of the Mycobacterium tuberculosis MscL (MtMscLΔC). A crystal structure of MtMscLΔC solubilized in the detergent n-dodecyl-β-D-maltopyranoside reveals the pentameric, closed state-like architecture for the membrane spanning region observed in the previously solved full-length MtMscL. Electrophysiological characterization demonstrates that MtMscLΔC retains mechano-sensitivity, but with conductance and tension sensitivity more closely resembling full length EcMscL than MtMscL. This study establishes that the C-terminal domain of MtMscL is not required for oligomerization of the full-length channel, but rather influences the tension sensitivity and conductance properties of the channel. The collective picture that emerges from these data is that each MscL channel structure has characteristic features, highlighting the importance of studying multiple homologs.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); Gordon and Betty Moore Foundation; Beckman Institute; National Institutes of Health (NIH); National Institute of General Medical Sciences (NIGMS); National Center for Research Resources (NCRR)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1624426

Journal Information:: Scientific Reports, Vol. 8, Issue 1; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

References (59)

Manipulating the permeation of charged compounds through the MscL nanovalve Yang, Li‐Min; Blount, Paul The FASEB Journal, Vol. 25, Issue 1 https://doi.org/10.1096/fj.10-170076	journal	September 2010
Refined Structure of the Porin from Rhodopseudomonas blastica Kreusch, Andreas; Schulz, Georg E. Journal of Molecular Biology, Vol. 243, Issue 5 https://doi.org/10.1006/jmbi.1994.1690	journal	November 1994
Structure and stability of the C-terminal helical bundle of the E. coli mechanosensitive channel of large conductance: Walton and Rees Walton, Troy A.; Rees, Douglas C. Protein Science, Vol. 22, Issue 11 https://doi.org/10.1002/pro.2360	journal	September 2013
A large-conductance mechanosensitive channel in E. coli encoded by mscL alone Sukharev, Sergei I.; Blount, Paul; Martinac, Boris Nature, Vol. 368, Issue 6468 https://doi.org/10.1038/368265a0	journal	March 1994
Crystal structures explain functional properties of two E. coli porins Cowan, S. W.; Schirmer, T.; Rummel, G. Nature, Vol. 358, Issue 6389 https://doi.org/10.1038/358727a0	journal	August 1992
PHENIX: a comprehensive Python-based system for macromolecular structure solution. Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.45787	text	January 2010
Structure of the MscL Homolog from Mycobacterium tuberculosis: A Gated Mechanosensitive Ion Channel Chang, G.; Spencer, R. H.; Lee, A. T. Science, Vol. 282, Issue 5397 https://doi.org/10.1126/science.282.5397.2220	journal	December 1998
Mechano-sensitization of mammalian neuronal networks through expression of the bacterial large-conductance mechanosensitive ion channel Soloperto, Alessandro; Boccaccio, Anna; Contestabile, Andrea Journal of Cell Science, Vol. 131, Issue 5 https://doi.org/10.1242/jcs.210393	journal	January 2018
MECHANOSENSITIVE CHANNELS OF ESCHERICHIA COLI :The MscL Gene, Protein, and Activities Sukharev, Sergei I.; Blount, Paul; Martinac, Boris Annual Review of Physiology, Vol. 59, Issue 1 https://doi.org/10.1146/annurev.physiol.59.1.633	journal	October 1997
The gating mechanism of the large mechanosensitive channel MscL Sukharev, Sergei; Betanzos, Monica; Chiang, Chien-Sung Nature, Vol. 409, Issue 6821 https://doi.org/10.1038/35055559	journal	February 2001
REFMAC 5 dictionary: organization of prior chemical knowledge and guidelines for its use Vagin, Alexei A.; Steiner, Roberto A.; Lebedev, Andrey A. Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12 https://doi.org/10.1107/S0907444904023510	journal	November 2004
Mechanosensitive Channels: What Can They Do and How Do They Do It? Haswell, Elizabeth S.; Phillips, Rob; Rees, Douglas C. Structure, Vol. 19, Issue 10 https://doi.org/10.1016/j.str.2011.09.005	journal	October 2011
Streptomycin potency is dependent on MscL channel expression Iscla, Irene; Wray, Robin; Wei, Shuguang Nature Communications, Vol. 5, Issue 1 https://doi.org/10.1038/ncomms5891	journal	September 2014
Functional and structural conservation in the mechanosensitive channel MscL implicates elements crucial for mechanosensation Moe, Paul C.; Blount, Paul; Kung, Ching Molecular Microbiology, Vol. 28, Issue 3 https://doi.org/10.1046/j.1365-2958.1998.00821.x	journal	April 1998
YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity Schumann, Ulrike; Edwards, Michelle D.; Rasmussen, Tim Proceedings of the National Academy of Sciences, Vol. 107, Issue 28 https://doi.org/10.1073/pnas.1001405107	journal	June 2010
A possible unifying principle for mechanosensation Kung, Ching Nature, Vol. 436, Issue 7051 https://doi.org/10.1038/nature03896	journal	August 2005
Structural Investigation of MscL Gating Using Experimental Data and Coarse Grained MD Simulations Deplazes, Evelyne; Louhivuori, Martti; Jayatilaka, Dylan PLoS Computational Biology, Vol. 8, Issue 9 https://doi.org/10.1371/journal.pcbi.1002683	journal	September 2012
A cytoplasmic helix is required for pentamer formation of the Escherichia coli MscL mechanosensitive channel Ando, Chie; Liu, Naili; Yoshimura, Kenjiro Journal of Biochemistry, Vol. 158, Issue 2 https://doi.org/10.1093/jb/mvv019	journal	February 2015
The Effect of Detergent, Temperature, and Lipid on the Oligomeric State of MscL Constructs: Insights from Mass Spectrometry Reading, Eamonn; Walton, Troy A.; Liko, Idlir Chemistry & Biology, Vol. 22, Issue 5 https://doi.org/10.1016/j.chembiol.2015.04.016	journal	May 2015
Cytoplasmic Domain of MscS Interacts with Cell Division Protein FtsZ: A Possible Non-Channel Function of the Mechanosensitive Channel in Escherichia Coli Koprowski, Piotr; Grajkowski, Wojciech; Balcerzak, Marcin PLOS ONE, Vol. 10, Issue 5 https://doi.org/10.1371/journal.pone.0127029	journal	May 2015
On the Conformation of the COOH-terminal Domain of the Large Mechanosensitive Channel MscL Anishkin, Andriy; Gendel, Vyacheslav; Sharifi, Neda A. Journal of General Physiology, Vol. 121, Issue 3 https://doi.org/10.1085/jgp.20028768	journal	February 2003
The Structure of an Open Form of an E. coli Mechanosensitive Channel at 3.45 A Resolution Wang, W.; Black, S. S.; Edwards, M. D. Science, Vol. 321, Issue 5893 https://doi.org/10.1126/science.1159262	journal	August 2008
S. aureus MscL Is a Pentamer In Vivo but of Variable Stoichiometries In Vitro: Implications for Detergent-Solubilized Membrane Proteins Dorwart, Michael R.; Wray, Robin; Brautigam, Chad A. PLoS Biology, Vol. 8, Issue 12 https://doi.org/10.1371/journal.pbio.1000555	journal	December 2010
MscL: channeling membrane tension Walton, Troy A.; Idigo, Chinenye A.; Herrera, Nadia Pflügers Archiv - European Journal of Physiology, Vol. 467, Issue 1 https://doi.org/10.1007/s00424-014-1535-x	journal	May 2014
Characterization of three novel mechanosensitive channel activities in Escherichia coli Edwards, Michelle D.; Black, Susan; Rasmussen, Tim Channels, Vol. 6, Issue 4 https://doi.org/10.4161/chan.20998	journal	July 2012
Mechanical coupling of the multiple structural elements of the large-conductance mechanosensitive channel during expansion Li, Jie; Guo, Jianli; Ou, Xiaomin Proceedings of the National Academy of Sciences, Vol. 112, Issue 34 https://doi.org/10.1073/pnas.1503202112	journal	August 2015
Generation and Evaluation of a Large Mutational Library from the Escherichia coli Mechanosensitive Channel of Large Conductance, MscL : IMPLICATIONS FOR CHANNEL GATING AND EVOLUTIONARY DESIGN Maurer, Joshua A.; Dougherty, Dennis A. Journal of Biological Chemistry, Vol. 278, Issue 23 https://doi.org/10.1074/jbc.M302892200	journal	April 2003
Structure of a tetrameric MscL in an expanded intermediate state Liu, Zhenfeng; Gandhi, Chris S.; Rees, Douglas C. Nature, Vol. 461, Issue 7260 https://doi.org/10.1038/nature08277	journal	August 2009
Defining the Physical Gate of a Mechanosensitive Channel, MscL, by Engineering Metal-Binding Sites Iscla, Irene; Levin, Gal; Wray, Robin Biophysical Journal, Vol. 87, Issue 5 https://doi.org/10.1529/biophysj.104.049833	journal	November 2004
Water Dynamics and Dewetting Transitions in the Small Mechanosensitive Channel MscS Anishkin, Andriy; Sukharev, Sergei Biophysical Journal, Vol. 86, Issue 5 https://doi.org/10.1016/S0006-3495(04)74340-4	journal	May 2004
Porin mutants with new channel properties Schmid, Benedikt; Maveyraud, Laurent; Krömer, Markus Protein Science, Vol. 7, Issue 7 https://doi.org/10.1002/pro.5560070714	journal	July 1998
Mechanosensitive Channels in Microbes Kung, Ching; Martinac, Boris; Sukharev, Sergei Annual Review of Microbiology, Vol. 64, Issue 1 https://doi.org/10.1146/annurev.micro.112408.134106	journal	October 2010
The oligomeric state of the truncated mechanosensitive channel of large conductance shows no variance in vivo: Oligomeric State of MscL: A Pentamer Iscla, Irene; Wray, Robin; Blount, Paul Protein Science, Vol. 20, Issue 9 https://doi.org/10.1002/pro.686	journal	July 2011
OCAM: A new tool for studying the oligomeric diversity of MscL channels: Oligomeric Diversity of MscL Channels Gandhi, Chris S.; Walton, Troy A.; Rees, Douglas C. Protein Science, Vol. 20, Issue 2 https://doi.org/10.1002/pro.562	journal	January 2011
Controlled delivery of bioactive molecules into live cells using the bacterial mechanosensitive channel MscL Doerner, Julia F.; Febvay, Sebastien; Clapham, David E. Nature Communications, Vol. 3, Issue 1 https://doi.org/10.1038/ncomms1999	journal	January 2012
Cavin1 intrinsically disordered domains are essential for fuzzy electrostatic interactions and caveola formation Tillu, Vikas A.; Rae, James; Gao, Ya Nature Communications, Vol. 12, Issue 1 https://doi.org/10.1038/s41467-021-21035-4	journal	February 2021
The crystal structure of mouse VDAC1 at 2.3 A resolution reveals mechanistic insights into metabolite gating Ujwal, R.; Cascio, D.; Colletier, J. -P. Proceedings of the National Academy of Sciences, Vol. 105, Issue 46 https://doi.org/10.1073/pnas.0809634105	journal	November 2008
Selectivity mechanism of the mechanosensitive channel MscS revealed by probing channel subconducting states Cox, C. D.; Nomura, T.; Ziegler, C. S. Nature Communications, Vol. 4, Issue 1 https://doi.org/10.1038/ncomms3137	journal	July 2013
PHENIX: a comprehensive Python-based system for macromolecular structure solution Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221 https://doi.org/10.1107/S0907444909052925	journal	January 2010
Single molecule FRET reveals pore size and opening mechanism of a mechano-sensitive ion channel Wang, Yong; Liu, Yanxin; DeBerg, Hannah A. eLife, Vol. 3 https://doi.org/10.7554/eLife.01834	journal	February 2014
Three Routes To Modulate the Pore Size of the MscL Channel/Nanovalve Yang, Li-Min; Wray, Robin; Parker, Juandell ACS Nano, Vol. 6, Issue 2 https://doi.org/10.1021/nn203703j	journal	January 2012
One face of a transmembrane helix is crucial in mechanosensitive channel gating Ou, X.; Blount, P.; Hoffman, R. J. Proceedings of the National Academy of Sciences, Vol. 95, Issue 19 https://doi.org/10.1073/pnas.95.19.11471	journal	September 1998
Towards an understanding of the structural and functional properties of MscL, a mechanosensitive channel in bacteria Blount, Paul; Sukharev, Sergei I.; Moe, Paul C. Biology of the Cell, Vol. 87, Issue 1-2 https://doi.org/10.1111/j.1768-322X.1996.tb00961.x	journal	January 1996
Microinjection Technique: Routine System for Characterization of Microcapillaries by Bubble Pressure Measurement Schnorf, Martin; Potrykus, Ingo; Neuhaus, Gunther Experimental Cell Research, Vol. 210, Issue 2 https://doi.org/10.1006/excr.1994.1038	journal	February 1994
Patch Clamp Electrophysiology for the Study of Bacterial Ion Channels in Giant Spheroplasts of E. coli Martinac, Boris; Rohde, Paul R.; Cranfield, Charles G. Methods in Molecular Biology https://doi.org/10.1007/978-1-62703-245-2_23	book	November 2012
Not Ions Alone: Barriers to Ion Permeation in Nanopores and Channels Beckstein, Oliver; Tai, Kaihsu; Sansom, Mark S. P. Journal of the American Chemical Society, Vol. 126, Issue 45 https://doi.org/10.1021/ja045271e	journal	November 2004
Pressure-sensitive ion channel in Escherichia coli. Martinac, B.; Buechner, M.; Delcour, A. H. Proceedings of the National Academy of Sciences, Vol. 84, Issue 8 https://doi.org/10.1073/pnas.84.8.2297	journal	April 1987
The Crystal Structure of a Five-Stranded Coiled Coil in COMP: A Prototype Ion Channel? Malashkevich, V. N.; Kammerer, R. A.; Efimov, V. P. Science, Vol. 274, Issue 5288 https://doi.org/10.1126/science.274.5288.761	journal	November 1996
Phaser crystallographic software McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D. Journal of Applied Crystallography, Vol. 40, Issue 4 https://doi.org/10.1107/S0021889807021206	journal	July 2007
Single residue substitutions that change the gating properties of a mechanosensitive channel in Escherichia coli. Blount, P.; Sukharev, S. I.; Schroeder, M. J. Proceedings of the National Academy of Sciences, Vol. 93, Issue 21 https://doi.org/10.1073/pnas.93.21.11652	journal	October 1996
Structural Models of the MscL Gating Mechanism Sukharev, Sergei; Durell, Stewart R.; Guy, H. Robert Biophysical Journal, Vol. 81, Issue 2 https://doi.org/10.1016/s0006-3495(01)75751-7	journal	August 2001
Purification of the Small Mechanosensitive Channel of Escherichia coli (MscS): the Subunit Structure, Conduction, and Gating Characteristicsin Liposomes Sukharev, Sergei Biophysical Journal, Vol. 83, Issue 1 https://doi.org/10.1016/s0006-3495(02)75169-2	journal	July 2002
Two types of mechanosensitive channels in the Escherichia coli cell envelope: solubilization and functional reconstitution Sukharev, S. I.; Martinac, B.; Arshavsky, V. Y. Biophysical Journal, Vol. 65, Issue 1 https://doi.org/10.1016/s0006-3495(93)81044-0	journal	July 1993
Estimation of the pore size of the large-conductance mechanosensitive ion channel of Escherichia coli Cruickshank, C. C.; Minchin, R. F.; Le Dain, A. C. Biophysical Journal, Vol. 73, Issue 4 https://doi.org/10.1016/s0006-3495(97)78223-7	journal	October 1997
A novel method for structure-based prediction of ion channel conductance properties Smart, O. S.; Breed, J.; Smith, G. R. Biophysical Journal, Vol. 72, Issue 3 https://doi.org/10.1016/s0006-3495(97)78760-5	journal	March 1997
Structural mechanism for gating of a eukaryotic mechanosensitive channel of small conductance Deng, Zengqin; Maksaev, Grigory; Schlegel, Angela M. Nature Communications, Vol. 11, Issue 1 https://doi.org/10.1038/s41467-020-17538-1	journal	July 2020
Correlating a Protein Structure with Function of a Bacterial Mechanosensitive Channel Moe, Paul C.; Levin, Gal; Blount, Paul Journal of Biological Chemistry, Vol. 275, Issue 40 https://doi.org/10.1074/jbc.m002971200	journal	June 2000
The MscS and MscL Families of Mechanosensitive Channels Act as Microbial Emergency Release Valves Booth, Ian R.; Blount, Paul Journal of Bacteriology, Vol. 194, Issue 18 https://doi.org/10.1128/jb.00576-12	journal	June 2012
Selectivity mechanisms in MscS-like channels Cox, Charles D.; Wann, Kenneth T.; Martinac, Boris Channels, Vol. 8, Issue 1 https://doi.org/10.4161/chan.27107	journal	November 2013

Cited By (1)

Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets Kapsalis, Charalampos; Wang, Bolin; El Mkami, Hassane Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-019-12591-x	journal	October 2019

Figures / Tables (9)

Similar Records

Force transduction and lipid binding in MscL: A continuum-molecular approach

Journal Article · Mon Dec 01 00:00:00 EST 2014 · PLoS ONE · OSTI ID:1624426

Vanegas, Juan M.; Arroyo, Marino; Fotiadis, Dimitrios

Local calcium signalling is mediated by mechanosensitive ion channels in mesenchymal stem cells

Journal Article · Sun Jan 22 00:00:00 EST 2017 · Biochemical and Biophysical Research Communications · OSTI ID:1624426

Vasileva, Valeria Y.; Pugovkina, Natalia A.; Vassilieva, Irina O.; +3 more

Crystal Structures of the Response Regulator DosR From Mycobacterium Tuberculosis Suggest a Helix Rearrangement Mechanism for Phosphorylation Activation

Journal Article · Tue May 26 00:00:00 EDT 2009 · J. Mol. Biol. 378:227,2008 · OSTI ID:1624426

Wisedchaisri, G; Wu, M; Sherman, D R; +1 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
MscL Channel
Sensitive Disposition
Permeation Pathway
MscL Homolog
Gold Sodium Thiosulfate
Permeation and transport
X-ray crystallography