Activation of Engineered Protein Tyrosine Phosphatases with the Biarsenical Compound AsCy3-EDT 2

• Published in: Chembiochem : a European journal of chemical biology 2017-10, Vol.18 (19), p.1950-1958
• Main Authors: Chan, Wai Cheung, Knowlton, Gregory S, Bishop, Anthony C
• Format: Article
• Language: English
• Subjects: Arsenicals - chemistry; Arsenicals - pharmacology; Dose-Response Relationship, Drug; Models, Molecular; Molecular Structure; Protein Engineering; Protein Tyrosine Phosphatases - chemistry; Protein Tyrosine Phosphatases - genetics; Protein Tyrosine Phosphatases - metabolism; Time Factors
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.201700253

Methods for activating signaling enzymes hold significant potential for the study of cellular signal transduction. Here we present a strategy for engineering chemically activatable protein tyrosine phosphatases (actPTPs). To generate actPTP1B, we introduced three cysteine point mutations in the enzyme's WPD loop. Biarsenical compounds were screened for the capability to bind actPTP1B's WPD loop and increase its phosphatase activity. We identified AsCy3-EDT as a robust activator that selectively targets actPTP1B in proteomic mixtures and intact cells. Introduction of the corresponding mutations in T-cell PTP also generates an enzyme (actTCPTP) that is strongly activated by AsCy3-EDT . Given the conservation of WPD-loop structure among the classical PTPs, our results potentially provide the groundwork of a widely generalizable approach for generating actPTPs as tools for elucidating PTP signaling roles as well as connections between dysregulated PTP activity and human disease.