Functional similarities between heterogeneously and homogenously expressed MscL constructs

详细信息    查看全文

• 作者：Gamma Chi ; Paul R. Rohde ; Pietro Ridone ; Ben Hankamer…
• 关键词：MscL ; Mechanosensitive channel ; Construct design ; Protein quantification ; Heterogeneous expression ; Patch clamp
• 刊名：European Biophysics Journal
• 出版年：2015
• 出版时间：October 2015
• 年：2015
• 卷：44
• 期：7
• 页码：589-598
• 全文大小：1,330 KB
• 参考文献：Ajouz B, Berrier C, Besnard M, Martinac B, Ghazi A (2000) Contributions of the different extramembranous domains of the mechanosensitive ion channel MscL to its response to membrane tension. J Biol Chem 275:1015鈥?022CrossRef PubMed
  Arkin IT, Sukharev SI, Blount P, Kung C, Brunger AT (1998) Helicity, membrane incorporation, orientation and thermal stability of the large conductance mechanosensitive ion channel from E. coli. Biochim Biophys Acta 1369:131鈥?40CrossRef PubMed
  Balleza D, Gomez-Lagunas F (2009) Conserved motifs in mechanosensitive channels MscL and MscS. Eur Biophys J 38:1013鈥?027CrossRef PubMed
  Bazykin GA, Kochetov AV (2011) Alternative translation start sites are conserved in eukaryotic genomes. Nucleic Acids Res 39:567鈥?77PubMed Central CrossRef PubMed
  Blount P, Sukharev SI, Schroeder MJ, Nagle SK, Kung C (1996) Single residue substitutions that change the gating properties of a mechanosensitive channel in Escherichia coli. Proc Natl Acad Sci USA 93:11652鈥?1657PubMed Central CrossRef PubMed
  Blount P, Schroeder MJ, Kung C (1997) Mutations in a bacterial mechanosensitive channel change the cellular response to osmotic stress. J Biol Chem 272:32150鈥?2157CrossRef PubMed
  Chang G, Spencer R, Lee A, Barclay M, Rees C (1998) Structure of the MscL homologue from Mycobacterium tuberculosis: a gated mechanosensitive ion channel. Science 282:2220鈥?226CrossRef PubMed
  Corry B, Rigby P, Liu ZW, Martinac B (2005) Conformational changes involved in MscL channel gating measured using FRET spectroscopy. Biophys J 89:L49鈥揕51PubMed Central CrossRef PubMed
  Corry B, Hurst AC, Pal P, Nomura T, Rigby P, Martinac B (2010) An improved open-channel structure of MscL determined from FRET confocal microscopy and simulation. J Gen Physiol 136:483鈥?94PubMed Central CrossRef PubMed
  Coste B, Xiao B, Santos JS, Syeda R, Grandl J, Spencer KS, Kim SE, Schmidt M, Mathur J, Dubin AE, Montal M, Patapoutian A (2012) Piezo proteins are pore-forming subunits of mechanically activated channels. Nature 483:176鈥?81PubMed Central CrossRef PubMed
  Delcour AH, Martinac B, Adler J, Kung C (1989) Modified reconstitution method used in patch-clamp studies of Escherichia coli ion channels. Biophys J 56:631鈥?36PubMed Central CrossRef PubMed
  Deplazes E, Louhivuori M, Jayatilaka D, Marrink SJ, Corry B (2012) Structural investigation of MscL gating using experimental data and coarse grained MD simulations. PLoS Comput Biol 8:e1002683PubMed Central CrossRef PubMed
  Doerner JF, Febvay S, Clapham DE (2012) Controlled delivery of bioactive molecules into live cells using the bacterial mechanosensitive channel MscL. Nat Commun 3:990PubMed Central CrossRef PubMed
  Foo A, Battle AR, Chi G, Hankamer B, Landsberg MJ, Martinac B (2015) Inducible release of particulates from liposomes using the mechanosensitive channel of large conductance and l -伪-lysophosphatidylcholine. doi:10.鈥?007/鈥媠00249-015-1055-4
  Frottin F, Martinez A, Peynot P, Mitra S, Holz RC, Giglione C, Meinnel T (2006) The proteomics of N-terminal methionine cleavage. Mol Cell Proteomics 5:2336鈥?349CrossRef PubMed
  Grage SL, Keleshian AM, Turdzeladze T, Battle AR, Tay WC, May RP, Holt SA, Contera SA, Haertlein M, Moulin M, Pal P, Rohde PR, Forsyth VT, Watts A, Huang KC, Ulrich AS, Martinac B (2011) Bilayer-mediated clustering and functional interaction of MscL channels. Biophys J 100:1252鈥?260PubMed Central CrossRef PubMed
  Harley CB, Reynolds RP (1987) Analysis of E. coli promoter sequences. Nucleic Acids Res 15:2343鈥?361PubMed Central CrossRef PubMed
  H盲se CC, Le Dain AC, Martinac B (1995) Purification and functional reconstitution of the recombinant large mechanosensitive ion channel (MscL) of Escherichia coli. J Biol Chem 270:18329鈥?8334CrossRef PubMed
  H盲se CC, Le Dain AC, Martinac B (1997a) Molecular dissection of the large mechanosensitive ion channel (MscL) of E. coli: mutants with altered channel gating and pressure sensitivity. J Membr Biol 157:17鈥?5CrossRef PubMed
  H盲se CC, Minchin RF, Kloda A, Martinac B (1997b) Cross-linking studies and membrane localization and assembly of radiolabelled large mechanosensitive ion channel (MscL) of Escherichia coli. Biochem Biophys Res Commun 232:777鈥?82CrossRef PubMed
  Hirel PH, Schmitter MJ, Dessen P, Fayat G, Blanquet S (1989) Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the side-chain length of the penultimate amino acid. Proce Natl Acad Sci USA 86:8247鈥?251CrossRef
  Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR (1989) Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51鈥?9CrossRef PubMed
  Iscla I, Wray R, Blount P (2008) On the structure of the N-terminal domain of the MscL channel: helical bundle or membrane interface. Biophys J 95:2283鈥?291PubMed Central CrossRef PubMed
  Kloda A, Ghazi A, Martinac B (2006) C-terminal charged cluster of MscL, RKKEE, functions as a pH sensor. Biophys J 90:1992鈥?998PubMed Central CrossRef PubMed
  Ko莽er A, Walko M, Feringa BL (2007) Synthesis and utilization of reversible and irreversible light-activated nanovalves derived from the channel protein MscL. Nat Protoc 2:1426鈥?437CrossRef PubMed
  Konijnenberg A, Yilmaz D, Ingolfsson HI, Dimitrova A, Marrink SJ, Li Z, Venien-Bryan C, Sobott F, Ko莽er A (2014) Global structural changes of an ion channel during its gating are followed by ion mobility mass spectrometry. Proc Natl Acad Sci USA 111:17170鈥?7175PubMed Central CrossRef PubMed
  Lee LM, Liu AP (2015) A microfluidic pipette array for mechanophenotyping of cancer cells and mechanical gating of mechanosensitive channels. Lab Chip 15:264鈥?73CrossRef PubMed
  Levina N, T枚temeyer S, Stokes NR, Louis P, Jones MA, Booth IR (1999) Protection of Escherichia coli cells against extreme turgor by activation of MscS and MscL mechanosensitive channels: identification of genes required for MscS activity. EMBO J 18:1730鈥?737PubMed Central CrossRef PubMed
  Ma J, Campbell A, Karlin S (2002) Correlations between Shine-Dalgarno sequences and gene features such as predicted expression levels and operon structures. J Bacteriol 184:5733鈥?745PubMed Central CrossRef PubMed
  Martinac B (2011) Bacterial mechanosensitive channels as a paradigm for mechanosensory transduction. Cell Physiol Biochem 28:1051鈥?060CrossRef PubMed
  Maurer JA, Dougherty DA (2001) A high-throughput screen for MscL channel activity and mutational phenotyping. Biochim Biophys Acta 1514:165鈥?69CrossRef PubMed
  Meyer GR, Gullingsrud J, Schulten K, Martinac B (2006) Molecular dynamics study of MscL interactions with a curved lipid bilayer. Biophys J 91:1630鈥?637PubMed Central CrossRef PubMed
  Park KH, Berrier C, Martinac B, Ghazi A (2004) Purification and functional reconstitution of N- and C-halves of the MscL channel. Biophys J 86:2129鈥?136PubMed Central CrossRef PubMed
  Perozo E, Kloda A, Cortes DM, Martinac B (2001) Site-directed spin-labeling analysis of reconstituted Mscl in the closed state. J Gen Physiol 118:193鈥?06PubMed Central CrossRef PubMed
  Perozo E, Cortes DM, Sompornpisut P, Kloda A, Martinac B (2002) Open channel structure of MscL and the gating mechanism of mechanosensitive channels. Nature 418:942鈥?48CrossRef PubMed
  Price CE, Ko莽er A, Kol S, van der Berg JP, Driessen AJ (2011) In vitro synthesis and oligomerization of the mechanosensitive channel of large conductance, MscL, into a functional ion channel. FEBS Lett 585:249鈥?54CrossRef PubMed
  Qi Y, Cheng X, Han W, Jo S, Schulten K, Im W (2014) CHARMM-GUI PACE CG builder for solution, micelle, and bilayer coarse-grained simulations. J Chem Inf Model 54:1003鈥?009PubMed Central CrossRef PubMed
  Saint N, Lacapere JJ, Gu LQ, Ghazi A, Martinac B, Rigaud JL (1998) A hexameric transmembrane pore revealed by two-dimensional crystallization of the large mechanosensitive ion channel (MscL) of Escherichia coli. J Biol Chem 273:14667鈥?4670CrossRef PubMed
  Sawada Y, Murase M, Sokabe M (2012) The gating mechanism of the bacterial mechanosensitive channel MscL revealed by molecular dynamics simulations: from tension sensing to channel opening. Channels (Austin, Tex) 6:317鈥?31CrossRef
  Sharif-Naeini R, Dedman A, Folgering JH, Duprat F, Patel A, Nilius B, Honore E (2008) TRP channels and mechanosensory transduction: insights into the arterial myogenic response. Pflugers Arch 456:529鈥?40CrossRef PubMed
  Stano M, Klucar L (2011) phiGENOME: an integrative navigation throughout bacteriophage genomes. Genomics 98:376鈥?80CrossRef PubMed
  Steinbacher S, Bass R, Strop P, Rees DC (2007) Structures of the prokaryotic mechanosensitive channels MscL and MscS. In: Hamill OP (ed) Current topics in membranes volume 58: mechanosensitive ion channels, Part A. Elsevier Inc, Amsterdam, pp 1鈥?4CrossRef
  Sukharev SI, Martinac B, Arshavsky VY, Kung C (1993) Two types of mechanosensitive channels in the Escherichia coli cell envelope: solubilization and functional reconstitution. Biophys J 65:177鈥?83PubMed Central CrossRef PubMed
  Sukharev SI, Blount P, Martinac B, Blattner FR, Kung C (1994) A large-conductance mechanosensitive channel in E. coli encoded by mscL alone. Nature 368:265鈥?68CrossRef PubMed
  Walton TA, Rees DC (2013) Structure and stability of the C-terminal helical bundle of the E. coli mechanosensitive channel of large conductance. Protein Sci 22:1592鈥?601PubMed Central CrossRef PubMed
  Wang Y, DeBerg H, Liu Y, Nomura T, Tonks-Hoffman M, Rohde PR, Schulten K, Martinac B, Selvin PR (2014) Single molecule FRET reveals pore size and opening mechanism of MscL. eLife 3:e01834PubMed Central PubMed
  Yang LM, Wray R, Parker J, Wilson D, Duran RS, Blount P (2012) Three routes to modulate the pore size of the MscL channel/nanovalve. ACS Nano 6:1134鈥?141PubMed Central CrossRef PubMed
  Yoshimura K, Batiza A, Schroeder M, Blount P, Kung C (1999) Hydrophilicity of a single residue within MscL correlates with increased channel mechanosensitivity. Biophys J 77:1960鈥?972PubMed Central CrossRef PubMed
  Yoshimura K, Batiza A, Kung C (2001) Chemically charging the pore constriction opens the mechanosensitive channel MscL. Biophys J 80:2198鈥?206PubMed Central CrossRef PubMed
  Yoshimura K, Usukura J, Sokabe M (2008) Gating-associated conformational changes in the mechanosensitive channel MscL. Proc Natl Acad Sci USA 105:4033鈥?038PubMed Central CrossRef PubMed
  
• 作者单位：Gamma Chi (1)
  Paul R. Rohde (2)
  Pietro Ridone (2)
  Ben Hankamer (1)
  Boris Martinac (2) (3)
  Michael J. Landsberg (1) (4)
  
  1. Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, 4072, Australia
  2. Molecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Darlinghurst, NSW, 2010, Australia
  3. St Vincent鈥檚 Clinical School, University of New South Wales, Darlinghurst, NSW, 2010, Australia
  4. School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, QLD, 4072, Australia
  
• 刊物类别：Physics and Astronomy
• 刊物主题：Physics
  Biophysics and Biomedical Physics
  Cell Biology
  Biochemistry
  Plant Physiology
  Animal Physiology
  Neurobiology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1017

文摘

The mechanosensitive channel of large conductance MscL is a well-characterized mechanically gated non-selective ion channel, which often serves as a prototype mechanosensitive channel for mechanotransduction studies. However, there are some discrepancies between MscL constructs used in these studies, most notably unintended heterogeneous expression from some MscL expression constructs. In this study we investigate the possible cause of this expression pattern, and compare the original non-homogenously expressing constructs with our new homogeneously expressing one to confirm that there is little functional difference between them. In addition, a new MscL construct has been developed with an improved molar extinction coefficient at 280 nm, enabling more accurate protein quantification. Keywords MscL Mechanosensitive channel Construct design Protein quantification Heterogeneous expression Patch clamp