Proteomics

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Clickable Glutathione Approach for Identification of Protein Glutathionylation in Response to Glucose Metabolism


ABSTRACT: Glucose metabolism and mitochondrial function are closely interconnected with cellular redox-homeostasis. Although glucose starvation, which often occurs in ischemic heart or insufficient vascularization of tumor cells, is known to induce redox-disturbance, 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 form 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 response to glucose starvation 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 1,300 proteins, including SMYD2, PP2Cα, and catalase. We further analyzed the cysteine modification site and functional implication of PP2Cα glutathionylation.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Eranthie Weerapana  

LAB HEAD: Eranthie Weerapana

PROVIDER: PXD004026 | Pride | 2018-10-26

REPOSITORIES: pride

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Publications

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

Samarasinghe Kusal T G KT   Munkanatta Godage Dhanushka N P DN   Zhou Yani Y   Ndombera Fidelis T FT   Weerapana Eranthie E   Ahn Young-Hoon YH  

Molecular bioSystems 20160701 8


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 mut  ...[more]

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