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Sensors 2017, 17(2), 315; doi:10.3390/s17020315

Depicting Binding-Mediated Translocation of HIV-1 Tat Peptides in Living Cells with Nanoscale Pens of Tat-Conjugated Quantum Dots

1,†
,
1,†,* and 2,†
1
The T.K.P. Research Center for Photonics, Chiao Tung University, Hsinchu 300, Taiwan
2
Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan 350, Taiwan
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Masateru Taniguchi
Received: 28 December 2016 / Revised: 24 January 2017 / Accepted: 30 January 2017 / Published: 10 February 2017
(This article belongs to the Special Issue Single-Molecule Sensing)
View Full-Text   |   Download PDF [4518 KB, uploaded 13 February 2017]   |  

Abstract

Cell-penetrating peptides (CPPs) can translocate across cell membranes, and thus have great potential for the cellular delivery of macromolecular cargoes. However, the mechanism of this cellular uptake process is not yet fully understood. In this study, a time-lapse single-particle light-sheet microscopy technique was implemented to obtain a parallel visualization of the translocating process of individual human immunodeficiency virus 1 (HIV-1) transactivator of transcription (Tat) peptide conjugated quantum dots (TatP-QDs) in complex cellular terrains. Here, TatP-QDs served as nanoscale dynamic pens, which depict remarkable trajectory aggregates of TatP-QDs on the cell surface. Spectral-embedding analysis of the trajectory aggregates revealed a manifold formed by isotropic diffusion and a fraction of directed movement, possibly caused by interaction between the Tat peptides and heparan sulfate groups on the plasma membrane. Further analysis indicated that the membrane deformation induced by Tat-peptide attachment increased with the disruption of the actin framework in cytochalasin D (cyto D)-treated cells, yielding higher interactions on the TatP-QDs. In native cells, the Tat peptides can remodel the actin framework to reduce their interaction with the local membrane environment. Characteristic hot spots for interaction were detected on the membrane, suggesting that a funnel passage may have formed for the Tat-coated particles. This finding offers valuable insight into the cellular delivery of nanoscale cargo, suggesting an avenue for direct therapeutic delivery. View Full-Text
Keywords: single-particle  tracking; cell-penetrating  peptides; heparan sulfate proteoglycan; actin filaments; live cell; plasma membrane single-particle  tracking; cell-penetrating  peptides; heparan sulfate proteoglycan; actin filaments; live cell; plasma membrane
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

  • Externally hosted supplementary file 1
    Link: https://zenodo.org/record/59990
    Description: All of the datasets and software used to support the conclusions of this article are available from the public data repository at the following website: https://zenodo.org/record/59990.

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MDPI and ACS Style

Lin, C.Y.; Huang, J.Y.; Lo, L.-W. Depicting Binding-Mediated Translocation of HIV-1 Tat Peptides in Living Cells with Nanoscale Pens of Tat-Conjugated Quantum Dots. Sensors 2017, 17, 315.

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