Relative Protein Lifetime Measurement in Plants Using Tandem Fluorescent Protein Timers

B - Book chapters etc edited externally

Zhang, H., Linster, E., Wirtz, M. and Theodoulou, F. L. 2022. Relative Protein Lifetime Measurement in Plants Using Tandem Fluorescent Protein Timers. in: Lois, L. M. and Trujillo, M. (ed.) Plant Proteostasis New York Humana Press Inc. pp. 201-220

AuthorsZhang, H., Linster, E., Wirtz, M. and Theodoulou, F. L.
EditorsLois, L. M. and Trujillo, M.
Abstract

Targeted protein degradation plays a wide range of important roles in plant growth and development, but analyzing protein turnover in vivo is technically challenging. Until recently, there has been no straightforward methodology for quantifying protein dynamics at subcellular resolution during cellular transitions in plants. A tandem fluorescent protein timer (tFT) is a fusion of two different fluorescent proteins with distinct fluorophore maturation kinetics, which allows estimation of relative protein age from the ratio of fluorescence intensities of the two fluorescent proteins. Here, we describe approaches to use this technology to report relative protein lifetime in both transient and stable plant transformation systems. tFTs enable in vivo, real-time protein lifetime assessment within subcellular compartments and across tissues, permitting the analysis of protein degradation dynamics in response to stresses or developmental cues and in different genetic backgrounds.

KeywordsProtein turnover; Fluorescent proteins; Confocal microscopy; Proteasome; GFP; mCherry
Page range201-220
Year of Publication2022
Book titlePlant Proteostasis
PublisherHumana Press Inc
Place of publicationNew York
Edition2nd
SeriesMethods in Molecular Biology, vol 2581
ISBNprint: 978-1-0716-2783-9
online: 978-1-0716-2784-6
ISSN1064-3745
Digital Object Identifier (DOI)https://doi.org/10.1007/978-1-0716-2784-6_14
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeTPM - Tailoring Plant Metabolism - Work package 1 (WP1) - High value lipids for health and industry
Open accessPublished as non-open access
Output statusPublished
Publication dates
Online23 Nov 2022

Permalink - https://repository.rothamsted.ac.uk/item/98qvw/relative-protein-lifetime-measurement-in-plants-using-tandem-fluorescent-protein-timers

Restricted files

Publisher's version

Under embargo indefinitely

17 total views
2 total downloads
5 views this month
0 downloads this month