Format

Send to

Choose Destination
J Am Chem Soc. 2015 Oct 14;137(40):12772-12775. doi: 10.1021/jacs.5b07875. Epub 2015 Oct 5.

Time-Resolved Proteomic Visualization of Dendrimer Cellular Entry and Trafficking.

Author information

1
Department of Biochemistry, Purdue University, West Lafayette, IN, 47907, United States.
2
Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, United States.
3
Department of Chemistry, Purdue University, West Lafayette, IN, 47907, United States.
4
Center for Cancer Research, Purdue University, West Lafayette, IN, 47907, United States.
#
Contributed equally

Abstract

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.

PMID:
26425924
PMCID:
PMC4766814
DOI:
10.1021/jacs.5b07875
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for American Chemical Society Icon for PubMed Central
Loading ...
Support Center