Access Type
Open Access Dissertation
Date of Award
January 2017
Degree Type
Dissertation
Degree Name
Ph.D.
Department
Chemistry
First Advisor
Young-Hoon Ahn
Abstract
Oxidative stress is a common characteristic of age-related diseases such as vascular diseases, diabetes and cancer. Many diseases are known to be regulated by glutathionylation. Glutathionylation is referred to as the formation of disulfide bond between a protein cysteine and a glutathione. To understand the molecular mechanisms behind the disease initiation and progression, identification of such glutathionylated proteins is important. Even though existing methods have been widely used, several limitations of these methods hinder the identification of such proteins in disease conditions. Therefore, we developed a versatile chemical method that generates clickable glutathione inside the cells. In this method, we generated a mutant form of glutathione synthetase (GS M4) to metabolically synthesize clickable glutathione for studying and identifying proteins in disease conditions. Using clickable glutathione approach, we uncovered a regulatory role of glucose metabolism in glutathionylation. Also, we have identified over 1400 of glutathionylated proteins in response to mitochondrial dysfunction and glucose deprivation, a common characteristic found in metabolic disorders. Moreover, we have identified several new glutathionylated proteins including ser/thr protein phosphatase 2Cα (PP2Cα). Further, we uncovered a new role for PP2Cα glutathionylation that promotes cancer cell migration through the activation of JNK in response to metabolic alterations.
Recommended Citation
Samarasinghe, Kusal Theekshana Gayan, "Development Of Chemical Tools To Investigate Protein S-Glutathionylation In Response To Metabolic Alteration" (2017). Wayne State University Dissertations. 1869.
https://digitalcommons.wayne.edu/oa_dissertations/1869