Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen
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Rabideau-2015-Translocation of non-canonical.pdf
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Author(s)
Rabideau, Amy E.
Liao, Xiaoli
Akcay, Gizem
Pentelute, Bradley L.
Date Issued
July 2015
Journal
Scientific Reports
Publisher
Nature Publishing Group
Citation
Rabideau, Amy E., Xiaoli Liao, Gizem Akcay, and Bradley L. Pentelute. “Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen.” Scientific Reports 5 (July 16, 2015): 11944.
Version
Final published version
Abstract
A variety of pathogenic bacteria infect host eukaryotic cells using protein toxins, which enter the cytosol and exert their cytotoxic effects. Anthrax lethal toxin, for example, utilizes the membrane-spanning translocase, protective antigen (PA) pore, to deliver the protein toxin lethal factor (LF) from the endosome into the cytosol of cells. Previous work has investigated the delivery of natural peptides and enzymatic domains appended to the C-terminus of the PA-binding domain of lethal factor (LF[subscript N]) into the cytosol via PA pore. Here, we move beyond natural amino acids and systematically investigate the translocation of polypeptide cargo containing non-canonical amino acids and functionalities through PA pore. Our results indicate translocation is not perturbed with alterations to the peptide backbone or side-chain. Moreover, despite their structural complexity, we found that the small molecule drugs, doxorubicin and monomethyl auristatin F (MMAF) translocated efficiently through PA pore. However, we found cyclic peptides and the small molecule drug docetaxel abrogated translocation due to their large size and structural rigidity. For cargos that reached the cytosol, we demonstrated that each remained intact after translocation. These studies show PA is capable of translocating non-canonical cargo provided it is in a conformational state conducive for passage through the narrow pore.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1038/srep11944
