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Brassinosteroids

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Cover of 'Brassinosteroids'

Table of Contents

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    Book Overview
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    Chapter 1 Protocol for Extraction and Isolation of Brassinosteroids from Plant Tissues
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    Chapter 2 Synthetic Protocol for AFCS: A Biologically Active Fluorescent Castasterone Analog Conjugated to an Alexa Fluor 647 Dye
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    Chapter 3 Physiological Analysis of Brassinosteroid Responses and Sensitivity in Rice
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    Chapter 4 Light Regulation of Brassinosteroid Signaling Components: Checking Regulation of Protein Stability in Darkness
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    Chapter 5 Approaches to Study Light Effects on Brassinosteroid Sensitivity
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    Chapter 6 A Technical Framework for Studying the Signaling Nexus of Brassinosteroids and Immunity
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    Chapter 7 Identification of Brassinosteroid Target Genes by Chromatin Immunoprecipitation Followed by High-Throughput Sequencing (ChIP-seq) and RNA-Sequencing
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    Chapter 8 Quantitation of Cell Type-Specific Responses to Brassinosteroid by Deep Sequencing of Polysome-Associated Polyadenylated RNA
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    Chapter 9 Methods for Modeling Brassinosteroid-Mediated Signaling in Plant Development
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    Chapter 10 Quantitative Microscopic Analysis of Plasma Membrane Receptor Dynamics in Living Plant Cells
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    Chapter 11 Analysis of In Vitro DNA Interactions of Brassinosteroid-Controlled Transcription Factors Using Electrophoretic Mobility Shift Assay
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    Chapter 12 Identification of Brassinosteroid Signaling Complexes by Coimmunoprecipitation and Mass Spectrometry
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    Chapter 13 Simplified Enrichment of Plasma Membrane Proteins from Arabidopsis thaliana Seedlings Using Differential Centrifugation and Brij-58 Treatment
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    Chapter 14 Probing Activation and Deactivation of the BRASSINOSTEROID INSENSITIVE1 Receptor Kinase by Immunoprecipitation
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    Chapter 15 The Primary Root of Sorghum bicolor (L. Moench) as a Model System to Study Brassinosteroid Signaling in Crops
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    Chapter 16 Brassinosteroid Action in Plant Abiotic Stress Tolerance
Attention for Chapter 12: Identification of Brassinosteroid Signaling Complexes by Coimmunoprecipitation and Mass Spectrometry
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Chapter title
Identification of Brassinosteroid Signaling Complexes by Coimmunoprecipitation and Mass Spectrometry
Chapter number 12
Book title
Brassinosteroids
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6813-8_12
Pubmed ID
Book ISBNs
978-1-4939-6811-4, 978-1-4939-6813-8
Authors

Walter van Dongen, Luc van Heerde, Sjef Boeren, Sacco C. de Vries

Editors

Eugenia Russinova, Ana I. Caño-Delgado

Abstract

A combination of coimmunoprecipitation (coIP) of tagged proteins followed by protein identification and quantitation using Liquid Chromatography Mass Spectrometry/Mass Spectrometry (LCMS/MS) has proven to be a reliable method to qualitatively characterize membrane-bound receptor complexes from plants. Success depends on a range of parameters, such as abundance and stability of the complex and functionality of the tagged receptors, efficiency of the protein complex isolation procedure, MS equipment, and analysis software in use. In this Chapter, we focus on the use of one of the green fluorescent protein-tagged receptors of the SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) family, of which SERK3, also known as BRASSINOSTEROID INSENSITIVE1 (BRI1) ASSOCIATED KINASE1 (BAK1), is a coreceptor of BRI1. Like BRI1 itself, SERK3 is a leucine-rich repeat receptor kinase (LRR RK) with a single-pass transmembrane domain. The latest updated laboratory protocol is presented as well as examples of data analysis and typical results obtained. Potential drawbacks of the procedure employed for plant membrane proteins will be pointed out.

Mendeley readers

The data shown below were compiled from readership statistics for 3 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 3 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 1 33%
Professor > Associate Professor 1 33%
Unknown 1 33%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 1 33%
Agricultural and Biological Sciences 1 33%
Unknown 1 33%