Assay to visualize specific protein oxidation reveals spatio-temporal regulation of SHP2

Nat Commun. 2017 Sep 6;8(1):466. doi: 10.1038/s41467-017-00503-w.


Reactive oxygen species are produced transiently in response to cell stimuli, and function as second messengers that oxidize target proteins. Protein-tyrosine phosphatases are important reactive oxygen species targets, whose oxidation results in rapid, reversible, catalytic inactivation. Despite increasing evidence for the importance of protein-tyrosine phosphatase oxidation in signal transduction, the cell biological details of reactive oxygen species-catalyzed protein-tyrosine phosphatase inactivation have remained largely unclear, due to our inability to visualize protein-tyrosine phosphatase oxidation in cells. By combining proximity ligation assay with chemical labeling of cysteine residues in the sulfenic acid state, we visualize oxidized Src homology 2 domain-containing protein-tyrosine phosphatase 2 (SHP2). We find that platelet-derived growth factor evokes transient oxidation on or close to RAB5+/ early endosome antigen 1- endosomes. SHP2 oxidation requires NADPH oxidases (NOXs), and oxidized SHP2 co-localizes with platelet-derived growth factor receptor and NOX1/4. Our data demonstrate spatially and temporally limited protein oxidation within cells, and suggest that platelet-derived growth factor-dependent "redoxosomes," contribute to proper signal transduction.Protein-tyrosine phosphatases (PTPs) are thought to be major targets of receptor-activated reactive oxygen species (ROS). Here the authors describe a method that allows the localized visualization of oxidized intermediates of PTPs inside cells during signaling, and provide support for the "redoxosome" model.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Cells, Cultured
  • Cysteine / genetics
  • Cysteine / metabolism
  • Hep G2 Cells
  • Humans
  • Immunoblotting
  • Mice
  • Mice, Knockout
  • Microscopy, Confocal / methods*
  • NADPH Oxidases / metabolism
  • Oxidation-Reduction / drug effects
  • Platelet-Derived Growth Factor / pharmacology
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / genetics
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / metabolism*
  • Reactive Oxygen Species / metabolism
  • Receptors, Platelet-Derived Growth Factor / metabolism
  • Signal Transduction*
  • Time-Lapse Imaging / methods*


  • Platelet-Derived Growth Factor
  • Reactive Oxygen Species
  • NADPH Oxidases
  • Receptors, Platelet-Derived Growth Factor
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Cysteine