FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells

Srinjan Basu; Lisa-Maria Needham; David Lando; Edward J. R. Taylor; Kai J. Wohlfahrt; Devina Shah; Wayne Boucher; Yi Lei Tan; Lawrence E. Bates; Olga Tkachenko; Julie Cramard; B. Christoffer Lagerholm; Christian Eggeling; Brian Hendrich; Dave Klenerman; Steven F. Lee; Ernest D. Laue

doi:10.1038/s41467-018-04486-0

Nature Communications (Jun 2018)

FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells

Srinjan Basu,
Lisa-Maria Needham,
David Lando,
Edward J. R. Taylor,
Kai J. Wohlfahrt,
Devina Shah,
Wayne Boucher,
Yi Lei Tan,
Lawrence E. Bates,
Olga Tkachenko,
Julie Cramard,
B. Christoffer Lagerholm,
Christian Eggeling,
Brian Hendrich,
Dave Klenerman,
Steven F. Lee,
Ernest D. Laue

Affiliations

Srinjan Basu: Department of Biochemistry, University of Cambridge
Lisa-Maria Needham: Department of Chemistry, University of Cambridge
David Lando: Department of Biochemistry, University of Cambridge
Edward J. R. Taylor: Department of Biochemistry, University of Cambridge
Kai J. Wohlfahrt: Department of Biochemistry, University of Cambridge
Devina Shah: Department of Biochemistry, University of Cambridge
Wayne Boucher: Department of Biochemistry, University of Cambridge
Yi Lei Tan: Department of Biochemistry, University of Cambridge
Lawrence E. Bates: Department of Biochemistry, University of Cambridge
Olga Tkachenko: Department of Biochemistry, University of Cambridge
Julie Cramard: Wellcome Trust – MRC Stem Cell Institute, University of Cambridge
B. Christoffer Lagerholm: Medical Research Council Human Immunology Unit and Wolfson Imaging Centre, Weatherall Institute of Molecular Medicine, University of Oxford
Christian Eggeling: Medical Research Council Human Immunology Unit and Wolfson Imaging Centre, Weatherall Institute of Molecular Medicine, University of Oxford
Brian Hendrich: Department of Biochemistry, University of Cambridge
Dave Klenerman: Department of Chemistry, University of Cambridge
Steven F. Lee: Department of Chemistry, University of Cambridge
Ernest D. Laue: Department of Biochemistry, University of Cambridge

DOI: https://doi.org/10.1038/s41467-018-04486-0
Journal volume & issue: Vol. 9, no. 1
pp. 1 – 14

Abstract

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Single molecule tracking of fluorescent proteins in live cells is temporally limited by fluorophore photobleaching. Here the authors show using fluorophore pairs that FRET competes with photobleaching to improve photostability and allow longer-term tracking of both single proteins and complexes.

Published in Nature Communications

ISSN: 2041-1723 (Online)
Publisher: Nature Portfolio
Country of publisher: United Kingdom
LCC subjects: Science
Website: https://www.nature.com/ncomms/

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