Abstract
Mechanistic understanding of biological and biochemical processes requires methods to analyze structures and states but also conformational dynamics of biomacromolecules at room temperature under physiologically relevant conditions. Single-molecule Förster resonance energy transfer (smFRET) has evolved to a versatile tool for exactly this, the observation of intra- and intermolecular conformational dynamics and interactions of biomacromolecules. We here outline the basic principles and illustrate the applications of smFRET for understanding molecular mechanisms of active membrane transporters.
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Literatur
Kalinin S, Peulen T, Sindbert S et al. (2012) A toolkit and benchmark study for FRET-restrained high-precision structural modeling. Nat Methods 9:1218–1225
Hohlbein J, Craggs TD, Cordes T (2014) Alternating-laser excitation: single-molecule FRET and beyond. Chem Soc Rev 43:1156–1171
Gouridis G, Schuurman-Wolters GK, Ploetz E et al. (2015) Conformational dynamics in substrate-binding domains influences transport in the ABC importer GlnPQ. Nat Struct Mol Biol 22:57–64
Lerner E, Cordes T, Ingargiola A et al. (2018) Toward dynamic structural biology: two decades of single-molecule Förster resonance energy transfer. Science 359:6373
Roy R, Hohng S, Ha T (2008) A practical guide to singlemolecule FRET. Nat Methods 5:507
Deniz AA, Dahan M, Grunwell JR et al. (1999) Single-pair fluorescence resonance energy transfer on freely diffusing molecules: observation of Förster distance dependence and subpopulations. Proc Natl Acad Sci USA 96:3670–3675
Ha T, Enderle T, Ogletree DF et al. (1996) Probing the interaction between two single molecules: fluorescence resonance energy transfer between a single donor and a single acceptor. Proc Natl Acad Sci USA 93:6264–6268
Hohng S, Lee S, Lee J et al. (2014) Maximizing information content of single-molecule FRET experiments: multi-color FRET and FRET combined with force or torque. Chem Soc Rev 43:1007–1013
Ploetz E, Lerner E, Husada F et al. (2016) Förster resonance energy transfer and protein-induced fluorescence enhancement as synergetic multi-scale molecular rulers. Sci Rep 6:33257
Craggs TD, Kapanidis AN (2012) Six steps closer to FRET-driven structural biology. Nat Methods 9:1157
Husada F, Gouridis G, Vietrov R et al. (2015) Watching conformational dynamics of ABC transporters with singlemolecule tools. Biochem Soc Transactions 43:1041
Locher KP (2016) Mechanistic diversity in ATP-binding cassette (ABC) transporters. Nat Struct Mol Biol 23:487–493
Liu Y, Liu Y, He L et al. (2018) Single-molecule fluorescence studies on the conformational change of the ABC transporter MsbA. Biophys Rep 4:153–165
Yang M, Levanon NL, Acar B et al. (2018) Single-molecule probing of the conformational homogeneity of the ABC transporter BtuCD. Nat Chem Biol 14:715–722
Husada FH, Bountra K, Tassis K et al. (2018) Conformational dynamics of the ABC transporter McjD seen by single-molecule FRET. In Revision.
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Rebecca Mächtel Jahrgang 1991. Biologiestudium an der Universität Erlangen-Nürnberg (B. Sc.) und TU München (M. Sc.), mit Aufenthalt an der Fondazione Edmund Mach, San Michele all‘Adige, Italien. Seit 2017 Promotion in Biologie an der LMU München.
Christian Gebhardt Jahrgang 1992. Physik- und Informatikstudium an der LMU München, mit Aufenthalt am Centre de Recherche Paul Pascal, Bordeaux, Frankreich. 2017 wissenschaftlicher Mitarbeiter am Max-Planck-Institut für Quantenoptik, Garching. Seit 2017 Promotion in Biologie an der LMU München.
Thorben Cordes Jahrgang 1980. Chemiestudium an der TU Braunschweig, mit Aufenthalt am University College Cork, Irland. 2008 Promotion in Physik an der LMU München, 2008–2009 Postdoc dort und 2010–2011 an der Universität Oxford, UK. 2011–2016 Assistant Professor, 2016–2017 Associate Professor an der Universität Groningen, Niederlande. Seit 2017 Professor für Physikalische und Synthetische Biologie an der LMU München.
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Mächtel, R., Gebhardt, C. & Cordes, T. Konformationsbewegungen von aktiven Membrantransportern. Biospektrum 24, 495–497 (2018). https://doi.org/10.1007/s12268-018-0945-2
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DOI: https://doi.org/10.1007/s12268-018-0945-2