title: Peroxide-dependent sulfenylation of the EGFR catalytic site enhances kinase activity.
abstract: Protein sulfenylation is a post-translational modification of emerging importance in higher eukaryotes. However, investigation of its diverse roles remains challenging, particularly within a native cellular environment. Herein we report the development and application of DYn-2, a new chemoselective probe for detecting sulfenylated proteins in human cells. These studies show that epidermal growth factor receptor-mediated signaling results in H(2)O(2) production and oxidation of downstream proteins. In addition, we demonstrate that DYn-2 has the ability to detect differences in sulfenylation rates within the cell, which are associated with differences in target protein localization. We also show that the direct modification of epidermal growth factor receptor by H(2)O(2) at a critical active site cysteine (Cys797) enhances its tyrosine kinase activity. Collectively, our findings reveal sulfenylation as a global signaling mechanism that is akin to phosphorylation and has regulatory implications for other receptor tyrosine kinases and irreversible inhibitors that target oxidant-sensitive cysteines in proteins.
authors.affiliation: Chemical Biology Graduate Program, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Candice E
authors.initials: CE
authors.lastname: Paulsen
authors.forename: Thu H
authors.initials: TH
authors.lastname: Truong
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Garcia
authors.forename: Arne
authors.initials: A
authors.lastname: Homann
authors.forename: Vinayak
authors.initials: V
authors.lastname: Gupta
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Leonard
authors.forename: Kate S
authors.initials: KS
authors.lastname: Carroll
mesh_terms: D020134:Catalytic Domain; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D021381:Protein Transport; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D015398:Signal Transduction; D013455:Sulfur / Q000378:metabolism*
chemical_list: D013455:Sulfur; D006861:Hydrogen Peroxide; D066246:ErbB Receptors; D017027:Protein Tyrosine Phosphatases