Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry

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

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

2 Citations (Scopus)

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.

LanguageEnglish
Title of host publicationBrassino-steroids
EditorsEugenia Russinova, Ana I. Caño-Delgado
PublisherHumana Press Inc.
Pages145-154
Number of pages10
ISBN (Electronic)9781493968138
ISBN (Print)9781493968114
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1564
ISSN (Print)1064-3745

Fingerprint

Brassinosteroids
Mass Spectrometry
Phosphotransferases
Plant Proteins
Proteins
Tandem Mass Spectrometry
Green Fluorescent Proteins
Leucine
Liquid Chromatography
Embryonic Development
Membrane Proteins
Software
Equipment and Supplies
Membranes

Keywords

  • Arabidopsis
  • GFP
  • Immunoprecipitation
  • Protein complex identification
  • Protein–protein interaction
  • Tandem mass spectrometry

Cite this

van Dongen, W., van Heerde, L., Boeren, S., & de Vries, S. C. (2017). Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry. In E. Russinova, & A. I. Caño-Delgado (Eds.), Brassino-steroids (pp. 145-154). (Methods in Molecular Biology; Vol. 1564). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6813-8_12
van Dongen, Walter ; van Heerde, Luc ; Boeren, Sjef ; de Vries, Sacco C. / Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry. Brassino-steroids. editor / Eugenia Russinova ; Ana I. Caño-Delgado. Humana Press Inc., 2017. pp. 145-154 (Methods in Molecular Biology).
@inbook{277813ef1dd645baa70c6b41a12ace01,
title = "Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry",
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.",
keywords = "Arabidopsis, GFP, Immunoprecipitation, Protein complex identification, Protein–protein interaction, Tandem mass spectrometry",
author = "{van Dongen}, Walter and {van Heerde}, Luc and Sjef Boeren and {de Vries}, {Sacco C.}",
year = "2017",
doi = "10.1007/978-1-4939-6813-8_12",
language = "English",
isbn = "9781493968114",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "145--154",
editor = "Eugenia Russinova and Ca{\~n}o-Delgado, {Ana I.}",
booktitle = "Brassino-steroids",

}

van Dongen, W, van Heerde, L, Boeren, S & de Vries, SC 2017, Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry. in E Russinova & AI Caño-Delgado (eds), Brassino-steroids. Methods in Molecular Biology, vol. 1564, Humana Press Inc., pp. 145-154. https://doi.org/10.1007/978-1-4939-6813-8_12

Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry. / van Dongen, Walter; van Heerde, Luc; Boeren, Sjef; de Vries, Sacco C.

Brassino-steroids. ed. / Eugenia Russinova; Ana I. Caño-Delgado. Humana Press Inc., 2017. p. 145-154 (Methods in Molecular Biology; Vol. 1564).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

TY - CHAP

T1 - Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry

AU - van Dongen, Walter

AU - van Heerde, Luc

AU - Boeren, Sjef

AU - de Vries, Sacco C.

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

KW - Arabidopsis

KW - GFP

KW - Immunoprecipitation

KW - Protein complex identification

KW - Protein–protein interaction

KW - Tandem mass spectrometry

U2 - 10.1007/978-1-4939-6813-8_12

DO - 10.1007/978-1-4939-6813-8_12

M3 - Chapter

SN - 9781493968114

T3 - Methods in Molecular Biology

SP - 145

EP - 154

BT - Brassino-steroids

A2 - Russinova, Eugenia

A2 - Caño-Delgado, Ana I.

PB - Humana Press Inc.

ER -

van Dongen W, van Heerde L, Boeren S, de Vries SC. Identification of brassinosteroid signaling complexes by coimmunoprecipitation and mass spectrometry. In Russinova E, Caño-Delgado AI, editors, Brassino-steroids. Humana Press Inc. 2017. p. 145-154. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6813-8_12