In vivo identification of plant protein complexes using IP-MS/MS

Jos R. Wendrich, Sjef Boeren, Barbara Möller, Dolf Weijers, Bert de Rybel

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

12 Citations (Scopus)

Abstract

Individual proteins often function as part of a protein complex. The identification of interacting proteins is therefore vital to understand the biological role and function of the studied protein. Here we describe a method for the in vivo identification of nuclear, cytoplasmic, and membrane-associated protein complexes from plant tissues using a strategy of immunoprecipitation followed by tandem mass spectrometry. By performing quantitative mass spectrometry measurements on biological triplicates, relative abundance of proteins in GFP-tagged complexes compared to background controls can be statistically evaluated to identify high-confidence interactors. We detail the entire workflow of this approach.

LanguageEnglish
Title of host publicationMethods in Molecular Biology
EditorsJürgen Kleine-Vehn, Michael Sauer
PublisherHumana Press Inc.
Pages147-158
Volume1497
ISBN (Electronic)9781493964697
ISBN (Print)9781493964673
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1497
ISSN (Print)1064-3745

Fingerprint

Plant Proteins
Proteins
Workflow
Nuclear Envelope
Tandem Mass Spectrometry
Immunoprecipitation
Mass Spectrometry
Membrane Proteins
Cell Membrane

Keywords

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

Cite this

Wendrich, J. R., Boeren, S., Möller, B., Weijers, D., & de Rybel, B. (2017). In vivo identification of plant protein complexes using IP-MS/MS. In J. Kleine-Vehn, & M. Sauer (Eds.), Methods in Molecular Biology (Vol. 1497, pp. 147-158). (Methods in Molecular Biology; Vol. 1497). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6469-7_14
Wendrich, Jos R. ; Boeren, Sjef ; Möller, Barbara ; Weijers, Dolf ; de Rybel, Bert. / In vivo identification of plant protein complexes using IP-MS/MS. Methods in Molecular Biology. editor / Jürgen Kleine-Vehn ; Michael Sauer. Vol. 1497 Humana Press Inc., 2017. pp. 147-158 (Methods in Molecular Biology).
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Wendrich, JR, Boeren, S, Möller, B, Weijers, D & de Rybel, B 2017, In vivo identification of plant protein complexes using IP-MS/MS. in J Kleine-Vehn & M Sauer (eds), Methods in Molecular Biology. vol. 1497, Methods in Molecular Biology, vol. 1497, Humana Press Inc., pp. 147-158. https://doi.org/10.1007/978-1-4939-6469-7_14

In vivo identification of plant protein complexes using IP-MS/MS. / Wendrich, Jos R.; Boeren, Sjef; Möller, Barbara; Weijers, Dolf; de Rybel, Bert.

Methods in Molecular Biology. ed. / Jürgen Kleine-Vehn; Michael Sauer. Vol. 1497 Humana Press Inc., 2017. p. 147-158 (Methods in Molecular Biology; Vol. 1497).

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

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T1 - In vivo identification of plant protein complexes using IP-MS/MS

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AB - Individual proteins often function as part of a protein complex. The identification of interacting proteins is therefore vital to understand the biological role and function of the studied protein. Here we describe a method for the in vivo identification of nuclear, cytoplasmic, and membrane-associated protein complexes from plant tissues using a strategy of immunoprecipitation followed by tandem mass spectrometry. By performing quantitative mass spectrometry measurements on biological triplicates, relative abundance of proteins in GFP-tagged complexes compared to background controls can be statistically evaluated to identify high-confidence interactors. We detail the entire workflow of this approach.

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KW - GFP

KW - Immunoprecipitation

KW - Protein complex identification

KW - Protein–protein interaction

KW - Tandem mass spectrometry

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Wendrich JR, Boeren S, Möller B, Weijers D, de Rybel B. In vivo identification of plant protein complexes using IP-MS/MS. In Kleine-Vehn J, Sauer M, editors, Methods in Molecular Biology. Vol. 1497. Humana Press Inc. 2017. p. 147-158. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6469-7_14