Abstract
Stress acclimation is initialized by sensing the stressor, transducing the signal, and inducing the response. In particular, the signal transduction is driven by protein–protein interactions and the response might involve de novo complex formation, shifts in subcellular localization and, thus, transportation that is mediated by other proteins. The investigation of protein–protein interactions and their regulation upon abiotic stress is crucial for a deeper understanding of the underlying mechanisms. FRET measurements by sensitized emission allow for the analysis of protein–protein interactions in real time and have a high potential to provide new insights into the regulation of protein–protein interaction with respect to subcellular localization and time. Within this section protocols are provided which allow for FRET analysis on the single cell level, the image acquisition procedure is described in detail and ImageJ plugins are suggested for the data evaluation.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Gadella T, van der Krogt G, Bisseling T (1999) GFP-based FRET microscopy in living plant cells. Trends Plant Sci 4:287–291
Müller SM, Galliardt H, Schneider J, Barisas B, Seidel T (2013) Quantification of Förster resonance energy transfer by monitoring sensitized emission in living plant cells. Front Plant Sci 4:413
Seidel T, Golldack D, Dietz KJ (2005) Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements. FEBS Lett 579:4374–4382
Seidel T, Schnitzer D, Golldack D, Dietz KJ (2008) Organelle-specific isoenzymes of plant V-ATPase as revealed by in vivo-FRET analysis. BMC Cell Biol 9:28
Schnitzer D, Seidel T, Sander T, Golldack D, Dietz KJ (2011) The cellular energization state affects peripheral stalk stability of plant vacuolar H+-ATPase and impairs vacuolar acidification. Plant Cell Physiol 52:946–956
Muthuramalingam M, Seidel T, Laxa M, de Miranda SM, Gärtner F, Wu JR, Ströher E, Kandlbinder A, Dietz KJ (2009) Multiple redox and non-redox interactions define 2-Cys peroxiredoxin as a regulatory hub in the chloroplast. Mol Plant 2:1273–1288
Klein P, Seidel T, Stöcker B, Dietz KJ (2012) The membrane-tethered transcription factor ANAC089 serves as redox-dependent regulator of stromal ascorbate peroxidase gene expression. Front Plant Sci 3:247
Seidel T, Seefeld B, Sauer M, Dietz KJ (2010) In vivo analysis of the 2-Cys peroxiredoxin by two-step FRET. J Biotechnol 149:272–279
Wolf H, Barisas BG, Dietz KJ, Seidel T (2013) Photo-convertible Kaede enables the detection of protein homo-aggregation and conformational alterations in plant cells. Mol Plant 6:1453–1462
Seefeldt B, Kasper R, Seidel T, Tinnefeld P, Dietz KJ, Heilemann M, Sauer M (2008) Fluorescent proteins for single-molecule fluorescence applications. J Biophotonics 1:74–82
Koushik SV, Chen H, Thaler C, Puhl HL, Vogel SS (2006) Cerulean, Venus, and VenusY67C FRET reference standards. Biophys J 91:L99–L101
Pandey S, Wang XQ, Coursol SA, Assmann SM (2001) Preparation and applications of Arabidopsis thaliana guard cell protoplasts. New Phytol 153:517–526
Beemiller P, Hoppe A, Swanson J (2006) A phosphatidylinositol-3-kinase-dependent signal transition regulates ARF1 and ARF6 during Fcγ receptor-mediated phagocytosis. PLoS Biol 4:e162
Hachet-Haas M, Converset N, Marchal O, Matthes H, Gioria S, Galzi J, Lecat S (2006) FRET co-localization analyzer—a method to validate measurements of sensitized emission FRET acquired by confocal microscopy and available as an ImageJ plug-in. Microsc Res Technnol 69:941–956
Youvan D, Silva C, Bylina E, Coleman W, Dilworth M, Yang M (1997) Calibration of fluorescence resonance energy transfer in microscopy using genetically engineered GFP-derivatives on nickel chelating beads. Biotechnology 3:1–18
Feige J, Sage D, Wahli W, Desvergne B, Gelman L (2005) PixFRET, an ImageJ plugin for FRET calculation that can accommodate variations in spectral bleed-throughs. Microsc Res Technol 68:51–58
Xia Z, Liu Y (2001) Reliable and global measurement of fluorescence resonance energy transfer using fluorescence microscopes. Biophys J 81:2395–3402
Roszik J, Lisboa D, Szollosi J, Vereb G (2009) Evaluation of intensity-based ratiometric FRET in image cytometry approaches and a software solution. Int Soc Adv Cytom A 74A:761–767
Seidel T, Kluge C, Hanitzsch M, Ross J, Sauer M, Dietz KJ, Golldack D (2004) Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells. J Biotechnol 112:165–175
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Seidel, T., Kirasi, D. (2017). Mining and Quantifying In Vivo Molecular Interactions in Abiotic Stress Acclimation. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 1631. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7136-7_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7136-7_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7134-3
Online ISBN: 978-1-4939-7136-7
eBook Packages: Springer Protocols