A Combinatorial Strategy for the Acquisition of Potent and Specific Protein Tyrosine Phosphatase Inhibitors

  • Sheng Zhang
  • Lan Chen
  • David S. Lawrence
  • Zhong-Yin ZhangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 928)


Protein tyrosine phosphatases (PTPs), a large family of signaling enzymes, regulate many cellular processes, such as proliferation, differentiation, migration, apoptosis, and immune responses. Small molecule inhibitors against PTPs are valuable both as powerful tools to study the functions of target PTPs and as lead compounds for pharmacological development. Here, we describe a novel combinatorial library approach to target simultaneously both the active site pocket and a peripheral secondary binding site for the acquisition of potent and specific PTP inhibitors. Fluorescence tagging during combinatorial library synthesis enables fluorescence polarization-based high-throughput screening of the resulting library, leading to identification of a TC-PTP inhibitor.

Key words

Protein tyrosine phosphatases (PTPs) PTP inhibitor Peripheral binding site Fluorescence tagged Combinatorial chemistry Fluorescence polarization High-throughput screening 



This work was supported by NIH grant RO1 CA126937, RO1 CA152194 RO1 CA69202, and RO1 CA79954.


  1. 1.
    Alonso A, Sasin J, Bottini N, Friedberg I, Osterman A, Godzik A, Hunter T, Dixon J, Mustelin T (2004) Protein tyrosine phosphatases in the human genome. Cell 117:699–711PubMedCrossRefGoogle Scholar
  2. 2.
    Tonks NK (2006) Protein tyrosine phosphatases: from genes, to function, to disease. Nat Rev Mol Cell Biol 7:833–846PubMedCrossRefGoogle Scholar
  3. 3.
    Zhang ZY (2001) Protein tyrosine phosphatases: prospects for therapeutics. Curr Opin Chem Biol 5:416–423PubMedCrossRefGoogle Scholar
  4. 4.
    Zhang ZY (2002) Protein tyrosine phosphatases: structure and function, substrate specificity, and inhibitor development. Annu Rev Pharmacol Toxicol 42:209–234PubMedCrossRefGoogle Scholar
  5. 5.
    Zhang ZY (2003) Chemical and mechanistic approaches to the study of protein tyrosine phosphatases. Acc Chem Res 36:385–392PubMedCrossRefGoogle Scholar
  6. 6.
    Shen K, Keng YF, Wu L, Guo XL, Lawrence DS, Zhang ZY (2001) Acquisition of a specific and potent PTP1B inhibitor from a novel combinatorial library and screening procedure. J Biol Chem 276:47311–47319PubMedCrossRefGoogle Scholar
  7. 7.
    Yu X, Sun JP, He Y, Guo X, Liu S, Zhou B, Hudmon A, Zhang ZY (2007) Structure, inhibitor, and regulatory mechanism of Lyp, a lymphoid-specific tyrosine phosphatase implicated in autoimmune diseases. Proc Natl Acad Sci USA 104:19767–19772PubMedCrossRefGoogle Scholar
  8. 8.
    Zhang S, Chen L, Luo Y, Gunawan A, Lawrence DS, Zhang ZY (2009) Acquisition of a potent and selective TC-PTP inhibitor via a stepwise fluorophore-tagged combinatorial synthesis and screening strategy. J Am Chem Soc 131:13072–13079PubMedCrossRefGoogle Scholar
  9. 9.
    Zhang X, He Y, Liu S, Yu Z, Jiang ZX, Yang Z, Dong Y, Nabinger SC, Wu L, Gunawan AM, Wang L, Chan RJ, Zhang ZY (2010) Salicylic acid based small molecule inhibitor for the oncogenic Src homology-2 domain containing protein tyrosine phosphatase-2 (SHP2). J Med Chem 53:2482–2493PubMedCrossRefGoogle Scholar
  10. 10.
    Zhou B, He Y, Zhang X, Xu J, Luo Y, Wang Y, Franzblau SG, Yang Z, Chan RJ, Liu Y, Zheng J, Zhang ZY (2010) Targeting mycobacterium protein tyrosine phosphatase B for antituberculosis agents. Proc Natl Acad Sci USA 107:4573–4578PubMedCrossRefGoogle Scholar
  11. 11.
    Zhang S, Chen L, Kumar S, Wu L, Lawrence DS, Zhang ZY (2007) An affinity-based fluorescence polarization assay for protein tyrosine phosphatases. Methods 42:261–267PubMedCrossRefGoogle Scholar
  12. 12.
    Roehrl MH, Wang JY, Wagner G (2004) A general framework for development and data analysis of competitive high-throughput screens for small-molecule inhibitors of protein-protein interactions by fluorescence polarization. Biochemistry 43:16056–16066PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Sheng Zhang
    • 1
  • Lan Chen
    • 2
  • David S. Lawrence
    • 3
  • Zhong-Yin Zhang
    • 1
    Email author
  1. 1.Department of Biochemistry and Molecular BiologyIndiana University School of MedicineIndianapolisUSA
  2. 2.Department of NeurologyIndiana University School of MedicineIndianapolisUSA
  3. 3.Division of Medicinal Chemistry and Natural Products, Department of ChemistryUniversity of North CarolinaChapel HillUSA

Personalised recommendations