Quantitative Tandem Affinity Purification, an Effective Tool to Investigate Protein Complex Composition in Plant Hormone Signaling: Strigolactones in the Spotlight

Frontiers in Plant Science
Sylwia StrukSofie Goormachtig

Abstract

Phytohormones tightly regulate plant growth by integrating changing environmental and developmental cues. Although the key players have been identified in many plant hormonal pathways, the molecular mechanisms and mode of action of perception and signaling remain incompletely resolved. Characterization of protein partners of known signaling components provides insight into the formed protein complexes, but, unless quantification is involved, does not deliver much, if any, information about the dynamics of the induced or disrupted protein complexes. Therefore, in proteomics research, the discovery of what actually triggers, regulates or interrupts the composition of protein complexes is gaining importance. Here, tandem affinity purification coupled to mass spectrometry (TAP-MS) is combined with label-free quantification (LFQ) to a highly valuable tool to detect physiologically relevant, dynamic protein-protein interactions in Arabidopsis thaliana cell cultures. To demonstrate its potential, we focus on the signaling pathway of one of the most recently discovered phytohormones, strigolactones.

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Citations

Aug 30, 2018·Plant, Cell & Environment·Sylwia StrukSofie Goormachtig
Feb 24, 2021·Emerging Topics in Life Sciences·Amanda L Smythers, Leslie M Hicks

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Datasets Mentioned

BETA
PXD009083

Methods Mentioned

BETA
tandem affinity purification
PCR
transgenic
electrophoresis
X-ray
acetylation
Two-Hybrid
Y2H
pull-down
co-immunoprecipitation

Software Mentioned

MaxQuant
Perseus
Andromeda
TAP
qTAP
MaxLFQ
LFQ

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