Glutathione Transferase Photoaffinity Labeling Displays GST Induction by Safeners And Pathogen Infection

Overview

Journal Plant Cell Physiol

Publisher Oxford University Press

Specialties Biology
Cell Biology

Date 2023 Nov 4

PMID 37924215

Authors

Maria Font Farre

Daniel Brown

Maurice Konig

Brian J Killinger

Farnusch Kaschani

Markus Kaiser

Aaron T Wright

Jonathan Burton

Renier A L van der Hoorn

Affiliations

Soon will be listed here.

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.

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