Endosomal binding kinetics of Eps15 and Hrs specifically regulate the degradation of RTKs
| Autor: | Frode Miltzow Skjeldal, Trygve Bergeland, Oddmund Bakke, Linda Hofstad Haugen |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Endosome lcsh:Medicine Endosomes Plasma protein binding Article Cell Line 03 medical and health sciences Humans Immunoprecipitation lcsh:Science Receptor Microscopy Confocal Multidisciplinary Endosomal Sorting Complexes Required for Transport Chemistry lcsh:R Autophosphorylation Receptor Protein-Tyrosine Kinases Signal transducing adaptor protein Fibroblasts Phosphoproteins Receptor–ligand kinetics Cell biology ESCRT complex Adaptor Proteins Vesicular Transport Kinetics Cytosol 030104 developmental biology lcsh:Q |
| Zdroj: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-13 (2017) |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-017-17320-2 |
| Popis: | Activation of EGF-R and PDGF-R triggers autophosphorylation and the recruitment of Eps15 and Hrs. These two endosomal proteins are important for specific receptor sorting. Hrs is recruiting ubiquitinated receptors to early endosomes to further facilitate degradation through the ESCRT complex. Upon receptor activation Hrs becomes phosphorylated and is relocated to the cytosol, important for receptor degradation. In this work we have studied the endosomal binding dynamics of Eps15 and Hrs upon EGF-R and PDGF-R stimulation. By analysing the fluorescence intensity on single endosomes after ligand stimulation we measured a time-specific decrease in the endosomal fluorescence level of Eps15-GFP and Hrs-YFP. Through FRAP experiments we could further register a specific change in the endosomal-membrane to cytosol binding properties of Eps15-GFP and Hrs-YFP. This specific change in membrane fractions proved to be a redistribution of the immobile fraction, which was not shown for the phosphorylation deficient mutants. We here describe a mechanism that can explain the previously observed relocation of Hrs from the endosomes to cytosol after EGF stimulation and show that Eps15 follows a similar mechanism. Moreover, this specific redistribution of the endosomal protein binding dynamics proved to be of major importance for receptor degradation. |
| Databáze: | OpenAIRE |
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