FRET-FLIM for visualizing and quantifying protein interactions in live plant cells

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

6 Citations (Scopus)

Abstract

Proteins are the workhorses that control most biological processes in living cells. Although proteins can accomplish their functions independently, the vast majority of functions require proteins to interact with other proteins or biomacromolecules. Protein interactions can be investigated through biochemical assays such as co-immunoprecipitation (co-IP) or Western blot analysis, but such assays lack spatial information. Here we describe a well-developed imaging method, Förster resonance energy transfer (FRET) analyzed by fluorescence lifetime imaging microscopy (FLIM), that can be used to visualize protein interactions with both spatial and temporal resolution in live cells. We demonstrate its use in plant developmental research by visualizing in vivo dimerization of AUXIN RESPONSE FACTOR (ARF) proteins, mediating auxin responses.

LanguageEnglish
Title of host publicationMethods in Molecular Biology
EditorsJürgen Kleine-Vehn, Michael Sauer
PublisherHumana Press Inc.
Pages135-146
Volume1497
ISBN (Electronic)9781493964697
ISBN (Print)9781493964673
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1497
ISSN (Print)1064-3745

Fingerprint

Optical Imaging
Energy Transfer
Plant Cells
Microscopy
Proteins
Biological Phenomena
Indoleacetic Acids
Dimerization
Immunoprecipitation
Western Blotting
Research

Keywords

  • FLIM
  • Fluorescent proteins
  • FRET
  • Protein interactions

Cite this

Rios, A. F., Radoeva, T., de Rybel, B., Weijers, D., & Borst, J. (2017). FRET-FLIM for visualizing and quantifying protein interactions in live plant cells. In J. Kleine-Vehn, & M. Sauer (Eds.), Methods in Molecular Biology (Vol. 1497, pp. 135-146). (Methods in Molecular Biology; Vol. 1497). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6469-7_13
Rios, Alejandra Freire ; Radoeva, Tatyana ; de Rybel, Bert ; Weijers, Dolf ; Borst, Janwillem. / FRET-FLIM for visualizing and quantifying protein interactions in live plant cells. Methods in Molecular Biology. editor / Jürgen Kleine-Vehn ; Michael Sauer. Vol. 1497 Humana Press Inc., 2017. pp. 135-146 (Methods in Molecular Biology).
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Rios, AF, Radoeva, T, de Rybel, B, Weijers, D & Borst, J 2017, FRET-FLIM for visualizing and quantifying protein interactions in live plant cells. in J Kleine-Vehn & M Sauer (eds), Methods in Molecular Biology. vol. 1497, Methods in Molecular Biology, vol. 1497, Humana Press Inc., pp. 135-146. https://doi.org/10.1007/978-1-4939-6469-7_13

FRET-FLIM for visualizing and quantifying protein interactions in live plant cells. / Rios, Alejandra Freire; Radoeva, Tatyana; de Rybel, Bert; Weijers, Dolf; Borst, Janwillem.

Methods in Molecular Biology. ed. / Jürgen Kleine-Vehn; Michael Sauer. Vol. 1497 Humana Press Inc., 2017. p. 135-146 (Methods in Molecular Biology; Vol. 1497).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

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AU - Rios, Alejandra Freire

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AU - Borst, Janwillem

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AB - Proteins are the workhorses that control most biological processes in living cells. Although proteins can accomplish their functions independently, the vast majority of functions require proteins to interact with other proteins or biomacromolecules. Protein interactions can be investigated through biochemical assays such as co-immunoprecipitation (co-IP) or Western blot analysis, but such assays lack spatial information. Here we describe a well-developed imaging method, Förster resonance energy transfer (FRET) analyzed by fluorescence lifetime imaging microscopy (FLIM), that can be used to visualize protein interactions with both spatial and temporal resolution in live cells. We demonstrate its use in plant developmental research by visualizing in vivo dimerization of AUXIN RESPONSE FACTOR (ARF) proteins, mediating auxin responses.

KW - FLIM

KW - Fluorescent proteins

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BT - Methods in Molecular Biology

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Rios AF, Radoeva T, de Rybel B, Weijers D, Borst J. FRET-FLIM for visualizing and quantifying protein interactions in live plant cells. In Kleine-Vehn J, Sauer M, editors, Methods in Molecular Biology. Vol. 1497. Humana Press Inc. 2017. p. 135-146. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6469-7_13