Elucidating the Role of Protein Sulfenylation in Eukaryotic Signal Transduction.

[摘要] H2O2 acts as a second messenger that can modulate intracellular signal transduction via chemoselective oxidation of cysteine residues in signaling proteins, however, the protein targets of H2O2 as well as how oxidation influences protein activity has remained largely unknown.In the present study, we present the first direct demonstration of an essential role for sulfenic acid modification of the thiol peroxidase Gpx3 to communicate conditions of oxidative stress to the transcription factor Yap1 in Saccharomyces cerevisiae.We then describe the characterization of methods to profile ROS production and to monitor the redox status of the cellular glutathione pool, as well as the development of a method to profile global protein sulfenylation in Saccharomyces cerevisiae.Next we present the development and application of an alkyne-based probe for sulfenic acids, DYn-2.This new probe was used to reveal dynamic changes in global protein sulfenylation in response to epidermal growth factor stimulation of the human epidermoid carcinoma A431 cell line.This study identified three protein tyrosine phosphatases and the epidermal growth factor receptor as direct protein targets of hydrogen peroxide produced for epidermal growth factor signaling.Additionally, oxidation of the epidermal growth factor receptor was shown to enhance inherent kinase activity.Collectively, these results provide novel insight into how hydrogen peroxide can function as a second messenger to regulate eukaryotic signaling pathways and have broad implications for therapeutic development.

[发布机构] University of Michigan

[效力级别] Redox Signaling

[关键词] Redox Biology;Redox Signaling;Protein Sulfenylation;Signal Transduction;Biological Chemistry;Science;Chemical Biology