Time-Resolved Proteomic Visualization of Dendrimer Cellular Entry and Trafficking
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- 作者：Linna Wang ; Li Yang ; Li Pan ; Naveen Reddy Kadasala ; Liang Xue ; Robert J. Schuster ; Laurie L. Parker ; Alexander Wei ; W. Andy Tao
- 刊名：Journal of the American Chemical Society
- 出版年：2015
- 出版时间：October 14, 2015
- 年：2015
- 卷：137
- 期：40
- 页码：12772-12775
- 全文大小：327K
- ISSN：1520-5126
文摘
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鈥揹endrimer 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.