Assay to visualize specific protein oxidation reveals spatio-temporal regulation of SHP2
| Autor: | Jana Harizanova, Philippe I. H. Bastiaens, Rabea Stockert, Katrin Schröder, Benjamin G. Neel, Ryouhei Tsutsumi |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Endosome Science Immunoblotting Phosphatase General Physics and Astronomy Protein Tyrosine Phosphatase Non-Receptor Type 11 Proximity ligation assay Protein tyrosine phosphatase Biology Protein oxidation Time-Lapse Imaging Article General Biochemistry Genetics and Molecular Biology Mice chemistry.chemical_compound 03 medical and health sciences Animals Humans Receptors Platelet-Derived Growth Factor ddc:610 Cysteine lcsh:Science Cells Cultured Mice Knockout Platelet-Derived Growth Factor chemistry.chemical_classification Reactive oxygen species Microscopy Confocal Multidisciplinary 030102 biochemistry & molecular biology NADPH Oxidases 3T3 Cells Hep G2 Cells General Chemistry Cell biology 030104 developmental biology chemistry Second messenger system lcsh:Q Sulfenic acid Signal transduction Reactive Oxygen Species Oxidation-Reduction Signal Transduction Proto-oncogene tyrosine-protein kinase Src |
| Zdroj: | Nature Communications Nature Communications, Vol 8, Iss 1, Pp 1-14 (2017) |
| ISSN: | 2041-1723 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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