Direct Chemical Activation of a Rationally Engineered Signaling Enzyme

Chio, Cynthia M.; Cheng, Karen W.; Bishop, Anthony C.

doi:10.1002/cbic.201500245

Cited by 4 publications

(3 citation statements)

References 43 publications

(37 reference statements)

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“…applied the “bump-hole” approach of allele-specific kinase inhibition to PTPs. They sensitized PTPs to biarsenical compounds creating via mutagenesis strategies non-natural allosteric-inhibition sites or changing the active sites, thereby controlling the function of each cellular phosphatase by designing selective mutant/inhibitor pairs. , Interestingly, recently they were able to apply that approach to create a SHP2 mutant that is activatable through binding of biarsenical compounds, showing the versatility of this approach.…”

Section: A Brief Guide To the Classification And Resources Of Phospha...mentioning

confidence: 99%

Approaches to Study Phosphatases

2016

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Phosphatases play key roles in normal physiology and diseases. Studying phosphatases has been both essential and challenging, and the application of conventional genetic and biochemical methods has led to crucial but still limited understanding of their mechanisms, substrates, and exclusive functions within highly intricate networks. With the advances in technologies such as cellular imaging and molecular and chemical biology in terms of sensitive tools and methods, the phosphatase field has thrived in the past years and has set new insights for cell signaling studies and for therapeutic development. In this review, we give an overview of the existing interdisciplinary tools for phosphatases, give examples on how they have been applied to increase our understanding of these enzymes, and suggest how they-and other tools yet barely used in the phosphatase field-might be adapted to address future questions and challenges.

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Section: A Brief Guide To the Classification And Resources Of Phospha...mentioning

confidence: 99%

Approaches to Study Phosphatases

2016

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“…Along these lines, we recently reported that a biarsenical compound (FlAsH-EDT 2 ) can directly activate a mutant of the PTP SHP2 through disruption of autoinhibitory interactions between SHP2’s N-terminal Src-homology 2 (SH2) domain and its PTP domain. [9] Unfortunately, the strategy developed for SHP2 activation is not broadly applicable across the PTP family, as only two of the 37 human classical PTPs utilize SH2-mediated autoinhibition. [4a] …”

Section: Introductionmentioning

confidence: 99%

Activation of Engineered Protein Tyrosine Phosphatases with the Biarsenical Compound AsCy3‐EDT₂

2017

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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-EDT2 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-EDT2. 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 misregulated PTP activity and human disease.

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“…By contrast, we have attempted to develop methods for engineering activatable PTPs that retain wild-type-like enzymatic activities and regulatory-control mechanisms until a small-molecule activator is administered 30,31 . In this vein, we recently reported that the phosphatase PTP1B can be rendered activatable by targeting the enzyme’s WPD loop, a structural feature that is conserved among classical PTPs 31 .…”

Section: Introductionmentioning

confidence: 99%

Chemical activation of divergent protein tyrosine phosphatase domains with cyanine-based biarsenicals

Plaman

Chan

Bishop

2019

Sci Rep

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Strategies for the direct chemical activation of specific signaling proteins could provide powerful tools for interrogating cellular signal transduction. However, targeted protein activation is chemically challenging, and few broadly applicable activation strategies for signaling enzymes have been developed. Here we report that classical protein tyrosine phosphatase (PTP) domains from multiple subfamilies can be systematically sensitized to target-specific activation by the cyanine-based biarsenical compounds AsCy3 and AsCy5. Engineering of the activatable PTPs (actPTPs) is achieved by the introduction of three cysteine residues within a conserved loop of the PTP domain, and the positions of the sensitizing mutations are readily identifiable from primary sequence alignments. In the current study we have generated and characterized actPTP domains from three different subfamilies of both receptor and non-receptor PTPs. Biarsenical-induced stimulation of the actPTPs is rapid and dose-dependent, and is operative with both purified enzymes and complex proteomic mixtures. Our results suggest that a substantial fraction of the classical PTP family will be compatible with the act-engineering approach, which provides a novel chemical-biological tool for the control of PTP activity and the study of PTP function.

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Direct Chemical Activation of a Rationally Engineered Signaling Enzyme

Cited by 4 publications

References 43 publications

Approaches to Study Phosphatases

Approaches to Study Phosphatases

Activation of Engineered Protein Tyrosine Phosphatases with the Biarsenical Compound AsCy3‐EDT₂

Chemical activation of divergent protein tyrosine phosphatase domains with cyanine-based biarsenicals

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Direct Chemical Activation of a Rationally Engineered Signaling Enzyme

Cited by 4 publications

References 43 publications

Approaches to Study Phosphatases

Approaches to Study Phosphatases

Activation of Engineered Protein Tyrosine Phosphatases with the Biarsenical Compound AsCy3‐EDT2

Chemical activation of divergent protein tyrosine phosphatase domains with cyanine-based biarsenicals

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Product

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Activation of Engineered Protein Tyrosine Phosphatases with the Biarsenical Compound AsCy3‐EDT₂