Targeting Protein-Protein Interactions of Tyrosine Phosphatases with Microarrayed Fragment Libraries Displayed on Phosphopeptide Substrate Scaffolds.

Authors:
Megan Hogan
Megan Hogan
Cold Spring Harbor Laboratory
Medhanit Bahta
Medhanit Bahta
Chemical Biology Laboratory
Boston | United States
Kohei Tsuji
Kohei Tsuji
Institute of Health Biosciences and Graduate School of Pharmaceutical Sciences
Scott Cherry
Scott Cherry
Macromolecular Crystallography Laboratory
George T Lountos
George T Lountos
Macromolecular Crystallography Laboratory
United States
Joseph E Tropea
Joseph E Tropea
National Cancer Institute
United States
Bryan M Zhao
Bryan M Zhao
McArdle Laboratory for Cancer Research

ACS Comb Sci 2019 Mar 31;21(3):158-170. Epub 2019 Jan 31.

Molecular and Translational Sciences Division , United States Army Medical Research Institute of Infectious Diseases , Frederick , Maryland 21702 , United States.

Chemical library screening approaches that focus exclusively on catalytic events may overlook unique effects of protein-protein interactions that can be exploited for development of specific inhibitors. Phosphotyrosyl (pTyr) residues embedded in peptide motifs comprise minimal recognition elements that determine the substrate specificity of protein tyrosine phosphatases (PTPases). We incorporated aminooxy-containing amino acid residues into a 7-residue epidermal growth factor receptor (EGFR) derived phosphotyrosine-containing peptide and subjected the peptides to solution-phase oxime diversification by reacting with aldehyde-bearing druglike functionalities. The pTyr residue remained unmodified. The resulting derivatized peptide library was printed in microarrays on nitrocellulose-coated glass surfaces for assessment of PTPase catalytic activity or on gold monolayers for analysis of kinetic interactions by surface plasmon resonance (SPR). Focusing on amino acid positions and chemical features, we first analyzed dephosphorylation of the peptide pTyr residues within the microarrayed library by the human dual-specificity phosphatases (DUSP) DUSP14 and DUSP22, as well as by PTPases from poxviruses (VH1) and Yersinia pestis (YopH). In order to identify the highest affinity oxime motifs, the binding interactions of the most active derivatized phosphopeptides were examined by SPR using noncatalytic PTPase mutants. On the basis of high-affinity oxime fragments identified by the two-step catalytic and SPR-based microarray screens, low-molecular-weight nonphosphate-containing peptides were designed to inhibit PTP catalysis at low micromolar concentrations.

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Source
http://pubs.acs.org/doi/10.1021/acscombsci.8b00122
Publisher Site
http://dx.doi.org/10.1021/acscombsci.8b00122DOI Listing
March 2019
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