Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5
Human α-defensin 5 (HD5) is a 32-residue cysteine-rich host-defense peptide that exhibits broad-spectrum antimicrobial activity and contributes to innate immunity in the human gut and other organ systems. Despite many years of investigation, its antimicrobial mechanism of action remains unclear. In...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | en_US |
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American Chemical Society (ACS)
2016
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| Online Access: | http://hdl.handle.net/1721.1/104065 https://orcid.org/0000-0002-2515-5901 https://orcid.org/0000-0002-6153-8803 https://orcid.org/0000-0002-5356-3638 |
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| author | Chileveru, Haritha Reddy Lim, Shion A. Chairatana, Phoom Wommack, Andrew Chiang, I-Ling Nolan, Elizabeth Marie |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Chileveru, Haritha Reddy Lim, Shion A. Chairatana, Phoom Wommack, Andrew Chiang, I-Ling Nolan, Elizabeth Marie |
| author_sort | Chileveru, Haritha Reddy |
| collection | MIT |
| description | Human α-defensin 5 (HD5) is a 32-residue cysteine-rich host-defense peptide that exhibits broad-spectrum antimicrobial activity and contributes to innate immunity in the human gut and other organ systems. Despite many years of investigation, its antimicrobial mechanism of action remains unclear. In this work, we report that HD5[subscript ox], the oxidized form of this peptide that exhibits three regiospecific disulfide bonds, causes distinct morphological changes to Escherichia coli and other Gram-negative microbes. These morphologies include bleb formation, cellular elongation, and clumping. The blebs are up to ∼1 μm wide and typically form at the site of cell division or cell poles. Studies with E. coli expressing cytoplasmic GFP reveal that HD5[subscript ox] treatment causes GFP emission to localize in the bleb. To probe the cellular uptake of HD5[subscript ox] and subsequent localization, we describe the design and characterization of a fluorophore–HD5 conjugate family. By employing these peptides, we demonstrate that fluorophore–HD5[subscript ox] conjugates harboring the rhodamine and coumarin fluorophores enter the E. coli cytoplasm. On the basis of the fluorescence profiles, each of these fluorophore–HD5[subscript ox] conjugates localizes to the site of cell division and cell poles. These studies support the notion that HD5[subscript ox'], at least in part, exerts its antibacterial activity against E. coli and other Gram-negative microbes in the cytoplasm. |
| first_indexed | 2024-09-23T13:09:09Z |
| format | Article |
| id | mit-1721.1/104065 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:09:09Z |
| publishDate | 2016 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1040652022-10-01T13:24:29Z Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 Chileveru, Haritha Reddy Lim, Shion A. Chairatana, Phoom Wommack, Andrew Chiang, I-Ling Nolan, Elizabeth Marie Massachusetts Institute of Technology. Department of Chemistry Nolan, Elizabeth Marie Chileveru, Haritha Reddy Lim, Shion A. Chairatana, Phoom Wommack, Andrew Chiang, I-Ling Nolan, Elizabeth Marie Human α-defensin 5 (HD5) is a 32-residue cysteine-rich host-defense peptide that exhibits broad-spectrum antimicrobial activity and contributes to innate immunity in the human gut and other organ systems. Despite many years of investigation, its antimicrobial mechanism of action remains unclear. In this work, we report that HD5[subscript ox], the oxidized form of this peptide that exhibits three regiospecific disulfide bonds, causes distinct morphological changes to Escherichia coli and other Gram-negative microbes. These morphologies include bleb formation, cellular elongation, and clumping. The blebs are up to ∼1 μm wide and typically form at the site of cell division or cell poles. Studies with E. coli expressing cytoplasmic GFP reveal that HD5[subscript ox] treatment causes GFP emission to localize in the bleb. To probe the cellular uptake of HD5[subscript ox] and subsequent localization, we describe the design and characterization of a fluorophore–HD5 conjugate family. By employing these peptides, we demonstrate that fluorophore–HD5[subscript ox] conjugates harboring the rhodamine and coumarin fluorophores enter the E. coli cytoplasm. On the basis of the fluorescence profiles, each of these fluorophore–HD5[subscript ox] conjugates localizes to the site of cell division and cell poles. These studies support the notion that HD5[subscript ox'], at least in part, exerts its antibacterial activity against E. coli and other Gram-negative microbes in the cytoplasm. United States. Army Research Office. Institute for Soldier Nanotechnologies (Contract W911NF-13-D-0001) National Science Foundation (U.S.) (Grant 007031) Massachusetts Institute of Technology (MIT UROP Program funds) Royal Thai Government (RTG) (Fellowship) Massachusetts Institute of Technology (2014 Richard R. Schrock summer graduate fellowship) National Institutes of Health (U.S.) (NIH Office of the Director, grant DP2OD007045) 2016-08-30T16:23:59Z 2016-08-30T16:23:59Z 2015-02 2015-02 Article http://purl.org/eprint/type/JournalArticle 0006-2960 1520-4995 http://hdl.handle.net/1721.1/104065 Chileveru, Haritha R., Shion A. Lim, Phoom Chairatana, Andrew J. Wommack, I-Ling Chiang, and Elizabeth M. Nolan. “ Visualizing Attack of Escherichia Coli by the Antimicrobial Peptide Human Defensin 5 .” Biochemistry 54, no. 9 (March 10, 2015): 1767-1777. https://orcid.org/0000-0002-2515-5901 https://orcid.org/0000-0002-6153-8803 https://orcid.org/0000-0002-5356-3638 en_US http://dx.doi.org/10.1021/bi501483q Biochemistry Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) Prof. Nolan via Erja Kajosalo |
| spellingShingle | Chileveru, Haritha Reddy Lim, Shion A. Chairatana, Phoom Wommack, Andrew Chiang, I-Ling Nolan, Elizabeth Marie Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 |
| title | Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 |
| title_full | Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 |
| title_fullStr | Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 |
| title_full_unstemmed | Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 |
| title_short | Visualizing Attack of Escherichia coli by the Antimicrobial Peptide Human Defensin 5 |
| title_sort | visualizing attack of escherichia coli by the antimicrobial peptide human defensin 5 |
| url | http://hdl.handle.net/1721.1/104065 https://orcid.org/0000-0002-2515-5901 https://orcid.org/0000-0002-6153-8803 https://orcid.org/0000-0002-5356-3638 |
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