Discovery and Characterization of Two Classes of Selective Inhibitors of the Suppressor of the TCR Signaling Family of Proteins

Weijie Zhou, Yue Yin, Emery Smith, Jacqueline Chou, Justin Shumate, Louis Scampavia, Timothy P. Spicer, Nicholas Carpino, Jarrod B. French

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The suppressor of T-cell receptor signaling (Sts) proteins, Sts-1, has recently emerged as a potential immunostimulatory target for drug development. Genetic inactivation of the Sts proteins dramatically increases host survival of systemic infection and leads to improved pathogen clearance. The protein tyrosine phosphatase (PTP) activity of these proteins arises from a C-terminal 2-histidine phosphatase (HP) domain. To identify new inhibitors of the HP activity of Sts-1, we miniaturized a phosphatase assay to a 1536-well format and conducted a 20 580 compound screen. Among the hits were two classes of structurally related compounds, tetracycline variants and sulfonated azo dyes. These hits had low micromolar to nanomolar IC 50 values. Orthogonal screening confirmed the validity of these inhibitors and demonstrated that both act competitively on Sts-1 phosphatase activity. When tested on other PTPs, PTP1B and SHP1, it was found that the tetracycline PTP1B, SHP1, the tetracycline variant (doxycycline), and the sulfonated azo dye (Congo red) are selective inhibitors of Sts-1 HP , with selectivity indices ranging from 19 to as high as 200. The planar polyaromatic moieties present in both classes of compounds suggested a common binding mode. The mutation of either tryptophan 494 or tyrosine 596, located near the active site of the protein, reduced the K i of the inhibitors from 3- to 18-fold, indicating that these residues may help to promote the binding of substrates with aromatic groups. This work provides new insights into substrate selectivity mechanisms and describes two classes of compounds that can serve as probes of function or as a basis for future drug discovery.

Original languageEnglish (US)
Pages (from-to)250-259
Number of pages10
JournalACS Infectious Diseases
Volume5
Issue number2
DOIs
StatePublished - Feb 8 2019
Externally publishedYes

Bibliographical note

Funding Information:
This work was supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health (NIH) through award number U01HL127522 (J.B.F., N.C.), the National Institute of Allergy and Infectious Disease of the NIH through award number R21AI130859 (J.B.F., N.C.), and the Office of the Assistant Secretary of Defense for Health Affairs through the Peer Reviewed Medical Research Program under award number W81XWH17-1-0147 (J.B.F., N.C.). In addition, parts of this work were supported by the National Institutes of General Medical Sciences of the NIH under grant number R35GM124898 (J.B.F.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or the Department of Defense. Additional support was provided by the Center for Biotechnology, a New York State Center for Advanced Technology, Stony Brook University, Cold Spring Harbor Laboratory, Brookhaven National Laboratory, and the Feinstein Institute for Medical Research.

Publisher Copyright:
© 2018 American Chemical Society.

Keywords

  • PTP
  • Sts
  • TULA
  • UBASH3
  • tetracycline

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