Quantifying degradation rates of transmembrane receptor kinases.

Details

Serval ID
serval:BIB_1E00B37A289B
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Quantifying degradation rates of transmembrane receptor kinases.
Journal
Methods in Molecular Biology
Author(s)
Geldner N.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Publication state
Published
Issued date
2011
Volume
779
Pages
217-224
Language
english
Abstract
Transmembrane receptor-kinases are widespread throughout eukaryotes and their activities are known to regulate all kinds of cellular responses in diverse organs and cell types. In order to guarantee the correct amplitude and duration of signals, receptor levels at the cellular surface need to be tightly controlled. The regulation of receptor degradation is the most direct way to achieve this and elaborate mechanisms are in place to control this process. Therefore, the rate of receptor degradation is a parameter of central importance for understanding the dynamics of a signal transduction cascade. Unfortunately, degradation of transmembrane receptors is a complicated multistep process that involves internalization from the plasma membrane, invagination into the lumen of endosomal compartments, and finally fusion with the vacuole for degradation by vacuolar proteases. Therefore, degradation should be measured in an as noninvasive way as possible, such as not to interfere with the complicated transport processes. Here, a method for minimally invasive, in vivo turn-over measurements in intact organs is provided. This technique was used for quantifying the turn-over rates of the Brassinosteroid receptor kinase BRI1 (BRASSINOSTEROID INSENSITIVE 1) in Arabidopsis thaliana root meristems. Pulse-chase expression of a fluorescently labeled BRI1 variant was used and its turn-over rate was determined by quantitative confocal microscopy. This method is well suited to measure turn-over of transmembrane kinases, but can evidently be extended to measure turn-over of any types of transmembrane proteins.
Keywords
Arabidopsis/enzymology, Arabidopsis/genetics, Arabidopsis Proteins/genetics, Arabidopsis Proteins/metabolism, Gene Expression Regulation, Plant, Hot Temperature, Membrane Proteins/genetics, Membrane Proteins/metabolism, Microscopy, Confocal/methods, Plants, Genetically Modified, Protein Kinases/genetics, Protein Kinases/metabolism, Proteolysis
Pubmed
Web of science
Create date
15/02/2012 15:59
Last modification date
20/08/2019 13:54
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