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Determination of Dynamics of Plant Plasma Membrane Proteins with Fluorescence Recovery and Raster Image Correlation Spectroscopy

SYSNO ASEP	0461678
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Determination of Dynamics of Plant Plasma Membrane Proteins with Fluorescence Recovery and Raster Image Correlation Spectroscopy
Author(s)	Laňková, Martina (UEB-Q)_{RID, ORCID} Humpolíčková, Jana (UFCH-W)_RID Vosolsobě, S. (CZ) Cit, Zdeněk (UEB-Q) Lacek, Jozef (UEB-Q)_ORCID Čovan, Martin (UEB-Q) Čovanová, Milada (UEB-Q)_{RID, ORCID} Hof, Martin (UFCH-W)_{RID, ORCID} Petrášek, Jan (UEB-Q)_{RID, ORCID}
Source Title	Microscopy and Microanalysis. - : Cambridge University Press - ISSN 1431-9276 Roč. 22, č. 2 (2016), s. 290-299
Number of pages	10 s.
Language	eng - English
Country	US - United States
Keywords	raster image correlation spectroscopy ; fluorescence recovery after photobleaching ; auxin influx
Subject RIV	EB - Genetics ; Molecular Biology
Subject RIV - cooperation	J. Heyrovsky Institute of Physical Chemistry - Physical ; Theoretical Chemistry
R&D Projects	GAP305/11/2476 GA ČR - Czech Science Foundation (CSF)
	GPP501/12/P951 GA ČR - Czech Science Foundation (CSF)
Institutional support	UEB-Q - RVO:61389030 ; UFCH-W - RVO:61388955
UT WOS	000378274600005
DOI	10.1017/S1431927616000568
Annotation	A number of fluorescence microscopy techniques are described to study dynamics of fluorescently labeled proteins, lipids, nucleic acids, and whole organelles. However, for studies of plant plasma membrane (PM) proteins, the number of these techniques is still limited because of the high complexity of processes that determine the dynamics of PM proteins and the existence of cell wall. Here, we report on the usage of raster image correlation spectroscopy (RICS) for studies of integral PM proteins in suspension-cultured tobacco cells and show its potential in comparison with the more widely used fluorescence recovery after photobleaching method. For RICS, a set of microscopy images is obtained by single-photon confocal laser scanning microscopy (CLSM). Fluorescence fluctuations are subsequently correlated between individual pixels and the information on protein mobility are extracted using a model that considers processes generating the fluctuations such as diffusion and chemical binding reactions. As we show here using an example of two integral PM transporters of the plant hormone auxin, RICS uncovered their distinct short-distance lateral mobility within the PM that is dependent on cytoskeleton and sterol composition of the PM. RICS, which is routinely accessible on modern CLSM instruments, thus represents a valuable approach for studies of dynamics of PM proteins in plants.
Workplace	Institute of Experimental Botany
Contact	David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469
Year of Publishing	2017

Number of the records: 1