Tandem Affinity Purification of Protein Complexes from Arabidopsis Cell Cultures

Marta García-León; Elisa Iniesto; Vicente Rubio

doi:10.1007/978-1-4939-7871-7_21

Tandem Affinity Purification of Protein Complexes from Arabidopsis Cell Cultures

Methods Mol Biol. 2018:1794:297-309. doi: 10.1007/978-1-4939-7871-7_21.

Authors

Marta García-León¹, Elisa Iniesto¹, Vicente Rubio²

Affiliations

¹ Departamento de Genética Molecular de Plantas, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
² Departamento de Genética Molecular de Plantas, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain. vrubio@cnb.csic.es.

PMID: 29855967
DOI: 10.1007/978-1-4939-7871-7_21

Abstract

Tandem affinity purification (TAP) coupled to mass spectrometry has become a powerful approach to identify protein-protein interactions from different biological systems, including plants, in a proteome-wide manner. By using two sequential affinity purification steps, TAP allows for isolation of high-purity TAP-tagged proteins of interest and their associated proteins. Here we describe optimized procedures to use the GS^Rhino TAP technology for protein complex isolation from Arabidopsis cell suspension cultures.

Keywords: Arabidopsis; Cell culture; Interactomics; Protein complex; Protein–protein interactions; Proteomics; Tandem affinity purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / metabolism*
Arabidopsis Proteins / isolation & purification*
Arabidopsis Proteins / metabolism*
Cells, Cultured
Protein Binding
Protein Interaction Mapping / methods*
Tandem Affinity Purification / methods*

Substances

Arabidopsis Proteins