Format

Send to

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

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

Author information

1
Laura and Isaac Perlmutter Cancer Center, NYU Langone Health, 430 East 29th Street, New York, NY, 10016, USA. Ryohei.Tsutsumi@nyumc.org.
2
Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Street 11, 44227, Dortmund, Germany.
3
Institute of Physiology, Goethe-University, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany.
4
Faculty of Chemistry and Chemical Biology, Technical University of Dortmund, Otto-Hahn Street 11, 44227, Dortmund, Germany.
5
Laura and Isaac Perlmutter Cancer Center, NYU Langone Health, 430 East 29th Street, New York, NY, 10016, USA.

Abstract

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.

PMID:
28878211
PMCID:
PMC5587708
DOI:
10.1038/s41467-017-00503-w
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center