Mass Spectrometry and Imaging Analysis of Nanoparticle-Containing Vesicles Provide a Mechanistic Insight into Cellular Trafficking
| Autor: | Markus B. Bannwarth, Volker Mailänder, Katharina Landfester, Simone-Franziska Glaser, Daniel Hofmann, Hansjörg Schild, Claudia Messerschmidt, Stefan Tenzer |
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| Rok vydání: | 2014 |
| Předmět: |
Proteomics
Endosome Vesicle Quantitative proteomics General Engineering General Physics and Astronomy Biological Transport Biology Endocytosis Mass Spectrometry Cell biology law.invention Microscopy Electron Transmission Confocal microscopy law Organelle Nanoparticles General Materials Science Nanocarriers Intracellular |
| Zdroj: | ACS Nano. 8:10077-10088 |
| ISSN: | 1936-086X 1936-0851 |
| DOI: | 10.1021/nn502754c |
| Popis: | Rational design of nanocarriers for drug delivery approaches requires an unbiased knowledge of uptake mechanisms and intracellular trafficking pathways. Here we dissected these processes using a quantitative proteomics approach. We isolated intracellular vesicles containing superparamagnetic iron oxide polystyrene nanoparticles and analyzed their protein composition by label-free quantitative mass spectrometry. The proteomic snapshot of organelle marker proteins revealed that an atypical macropinocytic-like mechanism mediated the entry of nanoparticles. We show that the entry mechanism is controlled by actin reorganization, atypical macropinocytic signaling, and ADP-ribosylation factor 1. Additionally, our proteomics data demonstrated a central role for multivesicular bodies and multilamellar lysosomes in trafficking and final nanoparticle storage. This was confirmed by confocal microscopy and cryo-TEM measurements. By quantitatively analyzing the protein composition of nanoparticle-containing vesicles, our study clearly defines the routes of nanoparticle entry, intracellular trafficking, and the proteomic milieu of a nanoparticle-containing vesicle. |
| Databáze: | OpenAIRE |
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