title: Time-Resolved Proteomic Visualization of Dendrimer Cellular Entry and Trafficking.
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.
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Linna
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Li
authors.initials: L
authors.lastname: Pan
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Naveen Reddy
authors.initials: NR
authors.lastname: Kadasala
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Liang
authors.initials: L
authors.lastname: Xue
authors.affiliation: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Schuster
authors.affiliation: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Laurie L
authors.initials: LL
authors.lastname: Parker
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN, 47907, United States.|Center for Cancer Research, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: Alexander
authors.initials: A
authors.lastname: Wei
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN, 47907, United States.|Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, United States.|Center for Cancer Research, Purdue University, West Lafayette, IN, 47907, United States.
authors.forename: W Andy
authors.initials: WA
authors.lastname: Tao
mesh_terms: D001692:Biological Transport; D050091:Dendrimers / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D040901:Proteomics
chemical_list: D050091:Dendrimers