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

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 enzy...

Full description

Saved in:
Bibliographic Details
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
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201700253