Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 137, Issue 40, Pages 12772-12775Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacs.5b07875
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Funding
- NIH [5R21RR025802, 5R01GM088317]
- Purdue University Center for Cancer Research [P30 CA023168]
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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 He La 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.
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