Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis
Abstract
The TAT protein transduction domain (PTD) has been used to deliver a wide variety of biologically active cargo for the treatment of multiple preclinical disease models, including cancer and stroke. However, the mechanism of transduction remains unknown. Because of the TAT PTD's strong cell-surface binding, early assumptions regarding cellular uptake suggested a direct penetration mechanism across the lipid bilayer by a temperature- and energy-independent process. Here we show, using a transducible TAT-Cre recombinase reporter assay on live cells, that after an initial ionic cell-surface interaction, TAT-fusion proteins are rapidly internalized by lipid raft-dependent macropinocytosis. Transduction was independent of interleukin-2 receptor/raft-, caveolar- and clathrin-mediated endocytosis and phagocytosis. Using this information, we developed a transducible, pH-sensitive, fusogenic dTAT-HA2 peptide that markedly enhanced TAT-Cre escape from macropinosomes. Taken together, these observations provide a scientific basis for the development of new, biologically active, transducible therapeutic molecules.
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TAT, HA2 and FHV peptides have previously been used to enhance the release of drugs from late-endosomes.
“These peptides have previously been successfully used to enhance the release of drugs from late-endosomes48,49.”
From GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain (2021) · cites this paper as [48]
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- Authors
- Wadia, J. S., Stan, R. V., & Dowdy, S. F.
- Affiliations
- Howard Hughes Medical Institute (US)
- University of California San Diego (US)
- Corresponding author
- Dowdy, Steven F
- Venue
- Nature Medicine
- Published
- 2004-02-08
- Volume
- 10
- Issue
- 3
- Pages
- 310-315
- ISSN
- 1078-8956 (ISSN-L)
- 1546-170X
- OpenAlex
- W2160863389
- Cited by
- 1,647
- References
- 1
- Citations by year
- 2026: 20 · 2025: 31 · 2024: 24 · 2023: 55 · 2022: 53 · 2021: 61 · 2020: 60 · 2019: 53 · 2018: 62 · 2017: 67 · 2016: 79 · 2015: 71 · 2014: 78 · 2013: 86 · 2012: 87
- FWCI
- 40.58
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Medicine (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Howard Hughes Medical Institute
- National Heart, Lung, and Blood Institute (HL065418, R01HL065418)
- National Cancer Institute (R01CA096098, CA96098)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advancements in Transdermal Drug Delivery
- Keywords
- TAT protein transduction domain, fusogenic peptides, TAT fusion protein, cell surface binding, pH-sensitive peptides, cellular uptake mechanisms, transduction mechanisms
- MeSH terms
- Amiloride, Animals, Biological Transport, Cell Line, Clathrin, Humans, Peptides, Pinocytosis, Endosomes, T-Lymphocytes, Transduction, Genetic, Viral Fusion Proteins, Cytochalasin D, Gene Products, tat, Nucleic Acid Synthesis Inhibitors, Membrane Microdomains, Caveolins, Caveolin 1, Mice
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