Time-Resolved Proteomic Visualization of Dendrimer Cellular Entry and Trafficking
| Autor: | Laurie L. Parker, Li Pan, Alexander Wei, Li Yang, Naveen Reddy Kadasala, W. Andy Tao, Linna Wang, Liang Xue, Robert J. Schuster |
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| Rok vydání: | 2015 |
| Předmět: |
Proteomics
Dendrimers Chemistry Systems biology media_common.quotation_subject Quantitative proteomics Endocytic cycle Nanotechnology Biological Transport General Chemistry Complex cell Endocytosis Biochemistry Catalysis Article Colloid and Surface Chemistry medicine.anatomical_structure Dendrimer medicine Biophysics Humans Internalization media_common HeLa Cells |
| Zdroj: | Journal of the American Chemical Society. 137(40) |
| ISSN: | 1520-5126 |
| Popis: | Our understanding of the complex cell entry pathways would greatly benefit from a comprehensive characterization of key proteins involved in this dynamic process. Here we devise a novel proteomic strategy named TITAN (Tracing Internalization and TrAfficking of Nanomaterials) to reveal real-time protein-dendrimer interactions using a systems biology approach. Dendrimers functionalized with photoreactive cross-linkers were internalized by HeLa cells and irradiated at set time intervals, then isolated and subjected to quantitative proteomics. In total, 809 interacting proteins cross-linked with dendrimers were determined by TITAN in a detailed temporal manner during dendrimer internalization, traceable to at least two major endocytic mechanisms, clathrin-mediated and caveolar/raft-mediated endocytosis. The direct involvement of the two pathways was further established by the inhibitory effect of dynasore on dendrimer uptake and changes in temporal profiles of key proteins. |
| Databáze: | OpenAIRE |
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