title: Cellular trajectories of peptide-modified gold particle complexes: comparison of nuclear localization signals and peptide transduction domains.
abstract: Gold nanoparticles modified with nuclear localization peptides were synthesized and evaluated for their subcellular distribution in HeLa human cervical epithelium cells, 3T3/NIH murine fibroblastoma cells, and HepG2 human hepatocarcinoma cells. Video-enhanced color differential interference contrast microscopy and transmission electron microscopy indicated that transport of nanoparticles into the cytoplasm and nucleus depends on peptide sequence and cell line. Recently, the ability of certain peptides, called protein transduction domains (PTDs), to transclocate cell and nuclear membranes in a receptor- and temperature-independent manner has been questioned (see for example, Lundberg, M.; Wikstrom, S.; Johansson, M. (2003) Mol. Ther. 8, 143-150). We have evaluated the cellular trajectory of gold nanoparticles carrying the PTD from HIV Tat protein. Our observations were that (1) the conjugates did not enter the nucleus of 3T3/NIH or HepG2 cells, and (2) cellular uptake of Tat PTD peptide-gold nanoparticle conjugates was temperature dependent, suggesting an endosomal pathway of uptake. Gold nanoparticles modified with the adenovirus nuclear localization signal and the integrin binding domain also entered cells via an energy-dependent mechanism, but in contrast to the Tat PTD, these signals triggered nuclear uptake of nanoparticles in HeLa and HepG2 cell lines.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Tkachenko
authors.forename: Huan
authors.initials: H
authors.lastname: Xie
authors.forename: Yanli
authors.initials: Y
authors.lastname: Liu
authors.forename: Donna
authors.initials: D
authors.lastname: Coleman
authors.forename: Joseph
authors.initials: J
authors.lastname: Ryan
authors.forename: Wilhelm R
authors.initials: WR
authors.lastname: Glomm
authors.forename: Mathew K
authors.initials: MK
authors.lastname: Shipton
authors.forename: Stefan
authors.initials: S
authors.lastname: Franzen
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Feldheim
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D015696:Gene Products, tat / Q000737:chemistry; D006046:Gold / Q000737:chemistry / Q000493:pharmacokinetics*; D006367:HeLa Cells; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D019913:Nuclear Localization Signals / Q000187:drug effects / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000493:pharmacokinetics*; D017434:Protein Structure, Tertiary / Q000502:physiology; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000493:pharmacokinetics; D013696:Temperature; D014161:Transduction, Genetic
chemical_list: D015696:Gene Products, tat; D046911:Macromolecular Substances; D019913:Nuclear Localization Signals; D010455:Peptides; D012710:Serum Albumin, Bovine; D006046:Gold