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|a 10.1093/jxb/erad418
|2 doi
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|a pubmed24n1482.xml
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|a (DE-627)NLM363709800
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|a (NLM)37878766
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
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|a Petutschnig, Elena Kristin
|e verfasserin
|4 aut
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|a A novel fluorescent protein pair facilitates FLIM-FRET analysis of plant immune receptor interaction under native conditions
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|c 2024
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
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|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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|a Date Completed 05.02.2024
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|a Date Revised 26.07.2024
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|a published: Print
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|a Citation Status MEDLINE
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|a © The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissionsoup.com.
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|a Elucidating protein-protein interactions is crucial for our understanding of molecular processes within living organisms. Microscopy-based techniques can detect protein-protein interactions in vivo at the single-cell level and provide information on their subcellular location. Fluorescence lifetime imaging microscopy (FLIM)-Förster resonance energy transfer (FRET) is one of the most robust imaging approaches, but it is still very challenging to apply this method to proteins which are expressed under native conditions. Here we describe a novel combination of fluorescence proteins (FPs), mCitrine and mScarlet-I, which is ideally suited for FLIM-FRET studies of low abundance proteins expressed from their native promoters in stably transformed plants. The donor mCitrine displays excellent brightness in planta, near-mono-exponential fluorescence decay, and a comparatively long fluorescence lifetime. Moreover, the FRET pair has a good spectral overlap and a large Förster radius. This allowed us to detect constitutive as well as ligand-induced interaction of the Arabidopsis chitin receptor components CERK1 and LYK5 in a set of proof-of-principle experiments. Due to the good brightness of the acceptor mScarlet-I, the FP combination can be readily utilized for co-localization studies. The FP pair is also suitable for co-immunoprecipitation experiments and western blotting, facilitating a multi-method approach for studying and confirming protein-protein interactions
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a in planta FLIM-FRET
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|a Arabidopsis
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|a Förster resonance energy transfer (FRET)
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|a co-immunoprecipitation
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| 650 |
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|a fluorescence lifetime imaging (FLIM)
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| 650 |
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|a mCitrine
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|a mScarlet-I
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| 650 |
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|a stably transformed plants
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|a Green Fluorescent Proteins
|2 NLM
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| 650 |
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|a 147336-22-9
|2 NLM
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| 700 |
1 |
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|a Pierdzig, Leon
|e verfasserin
|4 aut
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| 700 |
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|a Mittendorf, Josephine
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Niebisch, Jule Meret
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Lipka, Volker
|e verfasserin
|4 aut
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| 773 |
0 |
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|i Enthalten in
|t Journal of experimental botany
|d 1985
|g 75(2024), 3 vom: 02. Feb., Seite 746-759
|w (DE-627)NLM098182706
|x 1460-2431
|7 nnns
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| 773 |
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|g volume:75
|g year:2024
|g number:3
|g day:02
|g month:02
|g pages:746-759
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|u http://dx.doi.org/10.1093/jxb/erad418
|3 Volltext
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|d 75
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|e 3
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|h 746-759
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