Proximity labeling of protein complexes and cell type-specific organellar proteomes in Arabidopsis enabled by TurboID
- Stanford University, United States
- Carnegie Institution for Science, United States
Abstract
Defining specific protein interactions and spatially or temporally restricted local proteomes improves our understanding of all cellular processes, but obtaining such data is challenging, especially for rare proteins, cell types, or events. Proximity labeling enables discovery of protein neighborhoods defining functional complexes and/or organellar protein compositions. Recent technological improvements, namely two highly active biotin ligase variants (TurboID and miniTurbo), allowed us to address two challenging questions in plants: (1) what are in vivo partners of a low abundant key developmental transcription factor and (2) what is the nuclear proteome of a rare cell type? Proteins identified with FAMA-TurboID include known interactors of this stomatal transcription factor and novel proteins that could facilitate its activator and repressor functions. Directing TurboID to stomatal nuclei enabled purification of cell type- and subcellular compartment-specific proteins. Broad tests of TurboID and miniTurbo in Arabidopsis and N. benthamiana and versatile vectors enable customization by plant researchers.
Data availability
MS data have been depositedProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository (Vizcaino et al. 2013) and can be accessed through a reviewer account.Proximity labeling datasest:Dataset identifier: PXD013596FAMA-CFP AP-MS datasets:Dataset identifier: PXD013595
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Proximity labeling datasestProteomeXchange Consortium, PXD013596.
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FAMA-CFP AP-MS datasetsProteomeXchange Consortium, PXD013595.
Article and author information
Author details
Dominique C Bergmann
Funding
Howard Hughes Medical Institute
- Dominique C Bergmann
Austrian Science Fund (J4019-B29)
- Andrea Mair
National Institutes of Health (RO1-CA186568)
- Alice Y Ting
Carnegie Institution of Washington
- Shou-ling Xu
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2019, Mair et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Proximity labeling of protein complexes and cell type-specific organellar proteomes in Arabidopsis enabled by TurboID
eLife 8:e47864.
https://doi.org/10.7554/eLife.47864
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