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Glutathione transferase photoaffinity labeling displays GST induction by safeners and pathogen infection

Abstract:
Glutathione transferases (GSTs) represent a large and diverse enzyme family involved in the detoxification of small molecules by glutathione conjugation in crops, weeds and model plants. In this study, we introduce an easy and quick assay for photoaffinity labeling of GSTs to study GSTs globally in various plant species. The small-molecule probe contains glutathione, a photoreactive group and a minitag for coupling to reporter tags via click chemistry. Under UV irradiation, this probe quickly and robustly labels GSTs in crude protein extracts of different plant species. Purification and mass spectrometry (MS) analysis of labeled proteins from Arabidopsis identified 10 enriched GSTs from the Phi(F) and Tau(U) classes. Photoaffinity labeling of GSTs demonstrated GST induction in wheat seedlings upon treatment with safeners and in Arabidopsis leaves upon infection with avirulent bacteria. Treatment of Arabidopsis with salicylic acid (SA) analog benzothiadiazole (BTH) induces GST labeling independent of NPR1, the master regulator of SA. Six Phi- and Tau-class GSTs that are induced upon BTH treatment were identified, and their labeling was confirmed upon transient overexpression. These data demonstrate that GST photoaffinity labeling is a useful approach to studying GST induction in crude extracts of different plant species upon different types of stress.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1093/pcp/pcad132

Authors

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Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Chemistry
Sub department:
Chemistry Research Laboratory
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Role:
Author
ORCID:
0000-0002-3574-4347


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Funder identifier:
https://ror.org/0472cxd90
Grant:
101019324
616449


Publisher:
Oxford University Press
Journal:
Plant and Cell Physiology More from this journal
Volume:
65
Issue:
1
Pages:
128-141
Place of publication:
Japan
Publication date:
2023-10-31
Acceptance date:
2023-10-23
DOI:
EISSN:
1471-9053
ISSN:
0032-0781
Pmid:
37924215


Language:
English
Pubs id:
1560441
Local pid:
pubs:1560441
Deposit date:
2025-04-29
ARK identifier:

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