Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen
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)...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2015
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| Online Access: | http://hdl.handle.net/1721.1/98438 https://orcid.org/0000-0003-2659-7012 |
| _version_ | 1826198357732753408 |
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| author | Rabideau, Amy E. Liao, Xiaoli Akcay, Gizem Pentelute, Bradley L. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Rabideau, Amy E. Liao, Xiaoli Akcay, Gizem Pentelute, Bradley L. |
| author_sort | Rabideau, Amy E. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:03:35Z |
| format | Article |
| id | mit-1721.1/98438 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:03:35Z |
| publishDate | 2015 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/984382022-10-01T00:51:40Z Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen Rabideau, Amy E. Liao, Xiaoli Akcay, Gizem Pentelute, Bradley L. Massachusetts Institute of Technology. Department of Chemistry Rabideau, Amy E. Liao, Xiaoli Akcay, Gizem Pentelute, Bradley L. 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 Start-up Funds Massachusetts Institute of Technology. Charles E. Reed Faculty Initiative Fund Damon Runyon Cancer Research Foundation (Innovation Award) National Science Foundation (U.S.) (CAREER Award CHE-1351807) National Science Foundation (U.S.). Graduate Research Fellowship 2015-09-10T16:13:17Z 2015-09-10T16:13:17Z 2015-07 2014-10 Article http://purl.org/eprint/type/JournalArticle 2045-2322 http://hdl.handle.net/1721.1/98438 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. https://orcid.org/0000-0003-2659-7012 en_US http://dx.doi.org/10.1038/srep11944 Scientific Reports Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Nature Publishing Group Nature Publishing Group |
| spellingShingle | Rabideau, Amy E. Liao, Xiaoli Akcay, Gizem Pentelute, Bradley L. Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen |
| title | Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen |
| title_full | Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen |
| title_fullStr | Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen |
| title_full_unstemmed | Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen |
| title_short | Translocation of Non-Canonical Polypeptides into Cells Using Protective Antigen |
| title_sort | translocation of non canonical polypeptides into cells using protective antigen |
| url | http://hdl.handle.net/1721.1/98438 https://orcid.org/0000-0003-2659-7012 |
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