Journal article
Metabolic Synthesis of Clickable Glutathione for Chemoselective Detection of Glutathionylation
Journal of the American Chemical Society, v 136(33), pp 11566-11569
20 Aug 2014
PMID: 25079194
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Glutathionylation involves reversible protein cysteine modification that regulates the function of numerous proteins in response to redox stimuli, thereby altering cellular processes. Herein we developed a selective and versatile approach to identifying glutathionylation by using a mutant of glutathione synthetase (GS). GS wildtype catalyzes coupling of gamma Glu-Cys to Gly to form glutathione. We generated a GS mutant that catalyzes azido-Ala in place of Gly with high catalytic efficiency and selectivity. Transfection of this GS mutant (F152A/S151G) and incubation of azido-Ala in cells efficiently afford the azide-containing glutathione derivative, gamma Glu-Cys-azido-Ala. Upon H2O2 treatment, clickable glutathione allowed for selective and sensitive detection of glutathionylated proteins by Western blotting or fluorescence after click reaction with biotin-alkyne or rhodamine-alkyne. This approach affords the efficient metabolic tagging of intracellular glutathione with small clickable functionality, providing a versatile handle for characterizing glutathionylation.
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Details
- Title
- Metabolic Synthesis of Clickable Glutathione for Chemoselective Detection of Glutathionylation
- Creators
- Kusal T. G. Samarasinghe - Wayne State UniversityDhanushka N. P. Munkanatta Godage - Wayne State UniversityGarrett C. VanHecke - Wayne State UniversityYoung-Hoon Ahn - Wayne State University
- Publication Details
- Journal of the American Chemical Society, v 136(33), pp 11566-11569
- Publisher
- American Chemical Society; Washington, DC
- Number of pages
- 4
- Grant note
- WSU University Wayne State University
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- College of Arts and Sciences; Chemistry; Drexel University
- Web of Science ID
- WOS:000340737900002
- Scopus ID
- 2-s2.0-84906347101
- Other Identifier
- 991020099923904721
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- Web of Science research areas
- Chemistry, Multidisciplinary