Single-molecule fluorescence studies on the conformational change of the ABC transporter MsbA

Yanqing Liu; Yue Liu; Lingli He; Yongfang Zhao; Xuejun C. Zhang

doi:10.1007/s41048-018-0057-z

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Yanqing Liu, Yue Liu, Lingli He, Yongfang Zhao, Xuejun C. Zhang. Single-molecule fluorescence studies on the conformational change of the ABC transporter MsbA. Biophysics Reports, 2018, 4(3): 153-165. doi: 10.1007/s41048-018-0057-z

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Yanqing Liu, Yue Liu, Lingli He, Yongfang Zhao, Xuejun C. Zhang. Single-molecule fluorescence studies on the conformational change of the ABC transporter MsbA. Biophysics Reports, 2018, 4(3): 153-165. doi: 10.1007/s41048-018-0057-z

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Single-molecule fluorescence studies on the conformational change of the ABC transporter MsbA

doi: 10.1007/s41048-018-0057-z

1 National Laboratory of Biomacromolecules, National CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2 College of Life Science, University of Chinese Academy of Sciences, Beijing 100049, China

Funds: This study was supported by the National Basic Research Program of China (973 Program) (2014CB910400) and the National Natural Science Foundation of China (31522016 and 31470745).

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Corresponding author: Xuejun C. Zhang
Received Date: 02 April 2018
Rev Recd Date: 11 April 2018
Publish Date: 30 June 2018

Abstract

Abstract

ATP-binding cassette (ABC) transporters are found in all forms of life from microbes to humans, and transport a wide variety of substrates across the cell membrane using the energy released from ATP hydrolysis and an alternating-access mechanism. MsbA is a homodimeric ABC exporter from Gramnegative bacteria, and transports amphipathic substrates including precursors of lipopolysaccharides from the inner leaflet to the outer leaflet of the cytoplasmic membrane. Despite extensive structural and functional studies, controversies remain regarding the dynamic properties of the conformational changes of MsbA during its transport cycle in the lipid environment. Here, we used single-molecule fluorescence resonance energy transfer (smFRET) to explore the dynamic behaviors of MsbA in detergent micelles, nanodiscs, and proteoliposomes. MsbA reconstituted into liposomes showed higher transition frequency between different states on the cytoplasmic side, whereas detergent-solubilized MsbA showed higher transition frequency on the periplasmic side. Three major states were identified from this smFRET study in the functional cycle of MsbA, including an intermediate conformation between the fully opened and fully closed cytoplasmic conformations, associated with both ATP binding and hydrolysis.
- ABC transporter,
- Conformational change,
- Single-molecule fluorescence resonance energy transfer (smFRET)

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References(39)

References

Borbat PP, Surendhran K, Bortolus M, Zou P, Freed JH, Mchaourab HS (2007) Conformational motion of the ABC transporter MsbA induced by ATP hydrolysis. PLoS Biol 5:e271

Cooper RS, Altenberg GA (2013) Association/dissociation of the nucleotide-binding domains of the ATP-binding cassette protein MsbA measured during continuous hydrolysis. J Biol Chem 288:20785-20796

Dassa E (2011) Natural history of ABC systems:not only transporters. Essays Biochem 50:19-42

Davidson AL (2002) Mechanism of coupling of transport to hydrolysis in bacterial ATP-binding cassette transporters. J Bacteriol 184:1225-1233

Davidson AL, Chen J (2004) ATP-binding cassette transporters in bacteria. Annu Rev Biochem 73:241-268

Doerrler WT, Raetz CR (2002) ATPase activity of the MsbA lipid flippase of Escherichia coli. J Biol Chem 277:36697-36705

Doerrler WT, Reedy MC, Raetz CR (2001) An Escherichia coli mutant defective in lipid export. J Biol Chem 276:11461-11464

Doerrler WT, Gibbons HS, Raetz CR (2004) MsbA-dependent translocation of lipids across the inner membrane of Escherichia coli. J Biol Chem 279:45102-45109

Dong J, Yang G, McHaourab HS (2005) Structural basis of energy transduction in the transport cycle of MsbA. Science 308:1023-1028

Doshi R, Woebking B, van Veen HW (2010) Dissection of the conformational cycle of the multidrug/lipidA ABC exporter MsbA. Proteins 78:2867-2872

Eckford PD, Sharom FJ (2008) Functional characterization of Escherichia coli MsbA:interaction with nucleotides and substrates. J Biol Chem 283:12840-12850

Georgieva ER, Borbat PP, Ginter C, Freed JH, Boudker O (2013) Conformational ensemble of the sodium-coupled aspartate transporter. Nat Struct Mol Biol 20:215-221

Higgins CF (2007) Multiple molecular mechanisms for multidrug resistance transporters. Nature 446:749-757

Higgins CF, Linton KJ (2004) The ATP switch model for ABC transporters. Nat Struct Mol Biol 11:918-926

Jardetzky O (1966) Simple allosteric model for membrane pumps. Nature 211:969-970

Juette MF, Terry DS, Wasserman MR, Altman RB, Zhou Z, Zhao H, Blanchard SC (2016) Single-molecule imaging of nonequilibrium molecular ensembles on the millisecond timescale. Nat Methods 13:341-344

Lerner E, Cordes T, Ingargiola A, Alhadid Y, Chung SY, Michalet X, Weiss S (2018) Toward dynamic structural biology:two decades of single-molecule Forster resonance energy transfer. Science. https://doi.org/10.1126/science.aan1133

Locher KP (2016) Mechanistic diversity in ATP-binding cassette(ABC) transporters. Nat Struct Mol Biol 23:487-493

McKinney SA, Joo C, Ha T (2006) Analysis of single-molecule FRET trajectories using hidden markov modeling. Biophys J 91:1941-1951

Mi W, Li Y, Yoon SH, Ernst RK, Walz T, Liao M (2017) Structural basis of MsbA-mediated lipopolysaccharide transport. Nature 549:233-237

Moeller A, Lee SC, Tao H, Speir JA, Chang G, Urbatsch IL, Potter CS, Carragher B, Zhang Q (2015) Distinct conformational spectrum of homologous multidrug ABC transporters. Structure 23:450-460

Rigaud J-L, Pitard B, Levy D (1995) Reconstitution of membrane proteins into liposomes application to energy-transducing membrane proteins. Biochim Biophys Acta 1231:223-246

Ritchie TK, Grinkova YV, Bayburt TH, Denisov IG, Zolnerciks JK, Atkins WM, Sligar SG (2009) Reconstitution of membrane proteins in phospholipid bilayer nanodiscs. Methods Enzymol 464:211-231

Ruiz N, Kahne D, Silhavy TJ (2009) Transport of lipopolysaccharide across the cell envelope:the long road of discovery. Nat Rev Microbiol 7:677-683

Singh H, Velamakanni S, Deery MJ, Howard J, Wei SL, van Veen HW (2016) ATP-dependent substrate transport by the ABC transporter MsbA is proton-coupled. Nat Commun 7:12387

Smith CA, Rayment I (1996) X-ray structure of the magnesium(Ⅱ)ADP-vanadate complex of the dictyostelium discoideum Myosin Motor Domain to 1.9 Å resolution. Biochemistry 35:5404-5417

Urbatsch IL, Tyndall GA, Tombline G, Senior AE (2003) P-glycoprotein catalytic mechanism:studies of the ADP-vanadate inhibited state. J Biol Chem 278:23171-23179

Vafabakhsh R, Levitz J, Isacoff EY (2015) Conformational dynamics of a class C G-protein-coupled receptor. Nature 524:497-501

Venter H, Velamakanni S, Balakrishnan L, van Veen HW (2008) On the energy-dependence of Hoechst 33342 transport by the ABC transporter LmrA. Biochem Pharmacol 75:866-874

Ward A, Reyes CL, Yu J, Roth CB, Chang G (2007) Flexibility in the ABC transporter MsbA:alternating access with a twist. Proc Natl Acad Sci USA 104:19005-19010

Weng JW, Fan KN, Wang WN (2010) The conformational transition pathway of ATP binding cassette transporter MsbA revealed by atomistic simulations. J Biol Chem 285:3053-3063

Zhang XC, Han L, Zhao Y (2016) Thermodynamics of ABC transporters. Protein cell 7:17-27

Zhao Y, Quick M, Shi L, Mehler EL, Weinstein H, Javitch JA (2010a) Substrate-dependent proton antiport in neurotransmitter:sodium symporters. Nat Chem Biol 6:109-116

Zhao Y, Terry D, Shi L, Weinstein H, Blanchard SC, Javitch JA (2010b) Single-molecule dynamics of gating in a neurotransmitter transporter homologue. Nature 465:188-193

Zhao Y, Terry DS, Shi L, Quick M, Weinstein H, Blanchard SC, Javitch JA (2011) Substrate-modulated gating dynamics in a Na⁺-coupled neurotransmitter transporter homologue. Nature 474:109-113

Zhou Z, White KA, Polissi A, Georgopoulos C, Raetz CR (1998) Function of Escherichia coli MsbA, an essential ABC family transporter, in lipid A and phospholipid biosynthesis. J Biol Chem 273:12466-12475

Zhu Y, Zhang L, Zhang XC, Zhao Y (2017) Structural dynamics of Gia protein revealed by single molecule FRET. Biochem Biophys Res Commun 491:603-608

Zou P, Mchaourab HS (2010) Increased sensitivity and extended range of distance measurements in spin-labeled membrane proteins:Q-band double electron-electron resonance and nanoscale bilayers. Biophys J 98:L18-L20

Zou P, Bortolus M, McHaourab HS (2009) Conformational cycle of the ABC transporter MsbA in liposomes:detailed analysis using double electron-electron resonance spectroscopy. J Mol Biol 393:586-597

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