Identification of in planta protein–protein interactions using IP-MS

Suraj Jamge, Gerco Angenent, Marian Bemer*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

Gene regulation by transcription factors involves complex protein interaction networks, which include chromatin remodeling and modifying proteins as an integral part. Decoding these protein interactions is crucial for our understanding of chromatin-mediated gene regulation. Here, we describe a method for the immunoprecipitation of in planta nuclear protein complexes followed by mass spectrometry (IP-MS) to identify interactions between transcription factors and chromatin remodelers/modifiers in plants. In addition to a step-by-step bench protocol for immunoprecipitation and subsequent mass spectrometry, we provide guidelines and pointers on necessary controls and data analysis approaches.
Original languageEnglish
Title of host publicationPlant Chromatin Dynamics
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages315-329
ISBN (Electronic)9781493973187
ISBN (Print)9781493973170
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1675
ISSN (Print)1064-3745

Fingerprint

Immunoprecipitation
Chromatin
Mass Spectrometry
Transcription Factors
Protein Interaction Maps
Chromatin Assembly and Disassembly
Nuclear Proteins
Genes
Proteins
Guidelines

Keywords

  • Arabidopsis
  • Chromatin remodelers
  • Immunoprecipitation
  • Label free quantification
  • Mass spectrometry
  • Protein–protein interactions

Cite this

Jamge, S., Angenent, G., & Bemer, M. (2018). Identification of in planta protein–protein interactions using IP-MS. In Plant Chromatin Dynamics: Methods and Protocols (pp. 315-329). (Methods in Molecular Biology; Vol. 1675). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7318-7_18
Jamge, Suraj ; Angenent, Gerco ; Bemer, Marian. / Identification of in planta protein–protein interactions using IP-MS. Plant Chromatin Dynamics: Methods and Protocols. Humana Press Inc., 2018. pp. 315-329 (Methods in Molecular Biology).
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Jamge, S, Angenent, G & Bemer, M 2018, Identification of in planta protein–protein interactions using IP-MS. in Plant Chromatin Dynamics: Methods and Protocols. Methods in Molecular Biology, vol. 1675, Humana Press Inc., pp. 315-329. https://doi.org/10.1007/978-1-4939-7318-7_18

Identification of in planta protein–protein interactions using IP-MS. / Jamge, Suraj; Angenent, Gerco; Bemer, Marian.

Plant Chromatin Dynamics: Methods and Protocols. Humana Press Inc., 2018. p. 315-329 (Methods in Molecular Biology; Vol. 1675).

Research output: Chapter in Book/Report/Conference proceedingChapter

TY - CHAP

T1 - Identification of in planta protein–protein interactions using IP-MS

AU - Jamge, Suraj

AU - Angenent, Gerco

AU - Bemer, Marian

PY - 2018

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N2 - Gene regulation by transcription factors involves complex protein interaction networks, which include chromatin remodeling and modifying proteins as an integral part. Decoding these protein interactions is crucial for our understanding of chromatin-mediated gene regulation. Here, we describe a method for the immunoprecipitation of in planta nuclear protein complexes followed by mass spectrometry (IP-MS) to identify interactions between transcription factors and chromatin remodelers/modifiers in plants. In addition to a step-by-step bench protocol for immunoprecipitation and subsequent mass spectrometry, we provide guidelines and pointers on necessary controls and data analysis approaches.

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KW - Arabidopsis

KW - Chromatin remodelers

KW - Immunoprecipitation

KW - Label free quantification

KW - Mass spectrometry

KW - Protein–protein interactions

U2 - 10.1007/978-1-4939-7318-7_18

DO - 10.1007/978-1-4939-7318-7_18

M3 - Chapter

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EP - 329

BT - Plant Chromatin Dynamics

PB - Humana Press Inc.

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Jamge S, Angenent G, Bemer M. Identification of in planta protein–protein interactions using IP-MS. In Plant Chromatin Dynamics: Methods and Protocols. Humana Press Inc. 2018. p. 315-329. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7318-7_18