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A clickable glutathione approach for identification of protein glutathionylation in response to glucose metabolism
Journal article   Open access   Peer reviewed

A clickable glutathione approach for identification of protein glutathionylation in response to glucose metabolism

Kusal T G Samarasinghe, Dhanushka N P Munkanatta Godage, Yani Zhou, Fidelis T Ndombera, Eranthie Weerapana and Young-Hoon Ahn
Molecular bioSystems, v 12(8), pp 2471-2480
19 Jul 2016
DOI: https://doi.org/10.1039/c6mb00175k
PMID: 27216279
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://europepmc.org/articles/pmc4955733View
Accepted (AM)Open Access (License Unspecified),  Open

Abstract

Catalysis Catalytic Domain Gene Expression Glucose - metabolism Glutathione - metabolism Glutathione Synthase - chemistry Glutathione Synthase - genetics Glutathione Synthase - metabolism HEK293 Cells Humans Models, Molecular Mutation Oxidation-Reduction Protein Conformation Protein Interaction Domains and Motifs Protein Phosphatase 2C - chemistry Protein Phosphatase 2C - metabolism Protein Processing, Post-Translational Proteomics - methods Reactive Oxygen Species - metabolism
Glucose metabolism and mitochondrial function are closely interconnected with cellular redox-homeostasis. Although glucose starvation, which mimics ischemic conditions or insufficient vascularization, is known to perturb redox-homeostasis, global and individual protein glutathionylation in response to glucose metabolism or mitochondrial activity remains largely unknown. In this report, we use our clickable glutathione approach, which forms clickable glutathione (azido-glutathione) by using a mutant of glutathione synthetase (GS M4), for detection and identification of protein glutathionylation in response to glucose starvation. We found that protein glutathionylation is readily induced in HEK293 cells in response to low glucose concentrations when mitochondrial reactive oxygen species (ROS) are elevated in cells, and glucose is the major determinant for inducing reversible glutathionylation. Proteomic and biochemical analysis identified over 1300 proteins, including SMYD2, PP2Cα, and catalase. We further showed that PP2Cα is glutathionylated at C314 in a C-terminal domain, and PP2Cα C314 glutathionylation disrupts the interaction with mGluR3, an important glutamate receptor associated with synaptic plasticity.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemistry & Molecular Biology
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