Global analysis of protein tyrosine phosphatase activity with ultra-sensitive fluorescent probes

Sanjai Kumar, Bo Zhou, Fubo Liang, Heyi Yang, Wei Qing Wang, Zhong Yin Zhang

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Protein tyrosine phosphatases (PTPs) consist of a large family of enzymes known to play important roles in controlling virtually all aspects of cellular processes. However, assigning functional significance of PTPs in normal physiology and in diseases remains a major challenge in cell signaling. Since the function of a PTP is directly associated with its intrinsic activity, which is subject to post-translational regulation, new tools are needed to monitor the dynamic activities of PTPs, rather than mere abundance, on a global scale within the physiologically relevant environment of cells. To meet this objective, we report the synthesis and characterization of two rhodamine-conjugated probes that covalently label the active site of the PTPs in an activity-dependent manner, thus providing a direct readout of PTP activity and superior sensitivity, robustness, and quantifiability to previously reported biotinylated probes. We present evidence that the fluorescent probes can be used to identify new PTP markers and targets for potential diagnosis and treatment of human diseases. We also show that the fluorescent probes are capable of monitoring H2O2-mediated PTP inactivation, which should facilitate the study of regulated H2O2 production as a new tier of control over tyrosine phosphorylation-dependent signal transduction. The ability to profile the entire PTP family on the basis of changes in their activity is expected to yield new functional insights into pathways regulated by PTPs and contribute to the discovery of PTPs as novel therapeutic targets.

Original languageEnglish (US)
Pages (from-to)1898-1905
Number of pages8
JournalJournal of Proteome Research
Volume5
Issue number8
DOIs
StatePublished - Aug 1 2006

Keywords

  • Activity-based probes
  • Activity-based proteomics
  • Fluorescent probes
  • Proteomics
  • PTP
  • PTP activity profiling

ASJC Scopus subject areas

  • Genetics
  • Biotechnology
  • Biochemistry

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