Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells
Melittin is a membrane-active peptide with strong anticancer activity against various cancers. Despite decades of research, the role of the singular Trp in the anticancer activity and selectivity of melittin remains poorly understood. Here, we propose a theranostic solution based on the substitution...
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MDPI AG
2022-06-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/7/428 |
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| author | Yonghui Lv Xu Chen Zhidong Chen Zhanjun Shang Yongxiao Li Wanting Xu Yuan Mo Xinpei Wang Daiyun Xu Shengbin Li Zhe Wang Meiying Wu Junqing Wang |
| author_facet | Yonghui Lv Xu Chen Zhidong Chen Zhanjun Shang Yongxiao Li Wanting Xu Yuan Mo Xinpei Wang Daiyun Xu Shengbin Li Zhe Wang Meiying Wu Junqing Wang |
| author_sort | Yonghui Lv |
| collection | DOAJ |
| description | Melittin is a membrane-active peptide with strong anticancer activity against various cancers. Despite decades of research, the role of the singular Trp in the anticancer activity and selectivity of melittin remains poorly understood. Here, we propose a theranostic solution based on the substitution of Trp19 with a noncanonical fluorescent amino acid (Dap<sup>AMCA</sup>). The introduction of Dap<sup>AMCA</sup> residue in melittin stabilized the helical structure of the peptide, as evaluated by circular dichroism spectra and molecular dynamics simulations. In vitro hemolytic and anticancer activity assays revealed that introducing Dap<sup>AMCA</sup> residue in melittin changed its mode of action with the cell membrane, resulting in reduced hemolytic toxicity and an improved the selectivity index (SI), with up to a five-fold increase compared to melittin. In vitro fluorescence imaging of Dap<sup>AMCA</sup>-labeled melittin (MEL<sup>FL</sup>) in cancer cells demonstrated high membrane-penetrating activity, with strong nuclear and nucleolar localization ability. These findings provide implications for novel anticancer therapies based on Trp-substituted designs and nuclear/nucleolar targeted therapy. |
| first_indexed | 2024-03-09T12:54:36Z |
| format | Article |
| id | doaj.art-203f2f35e84c4b73ab82c03f902a5afd |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T12:54:36Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-203f2f35e84c4b73ab82c03f902a5afd2023-11-30T22:01:08ZengMDPI AGToxins2072-66512022-06-0114742810.3390/toxins14070428Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor CellsYonghui Lv0Xu Chen1Zhidong Chen2Zhanjun Shang3Yongxiao Li4Wanting Xu5Yuan Mo6Xinpei Wang7Daiyun Xu8Shengbin Li9Zhe Wang10Meiying Wu11Junqing Wang12School of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaDepartment of Pathology, The Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen 518033, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaSchool of Pharmaceutical Sciences, Shenzhen Campus of Sun Yat-Sen University, Shenzhen 518107, ChinaMelittin is a membrane-active peptide with strong anticancer activity against various cancers. Despite decades of research, the role of the singular Trp in the anticancer activity and selectivity of melittin remains poorly understood. Here, we propose a theranostic solution based on the substitution of Trp19 with a noncanonical fluorescent amino acid (Dap<sup>AMCA</sup>). The introduction of Dap<sup>AMCA</sup> residue in melittin stabilized the helical structure of the peptide, as evaluated by circular dichroism spectra and molecular dynamics simulations. In vitro hemolytic and anticancer activity assays revealed that introducing Dap<sup>AMCA</sup> residue in melittin changed its mode of action with the cell membrane, resulting in reduced hemolytic toxicity and an improved the selectivity index (SI), with up to a five-fold increase compared to melittin. In vitro fluorescence imaging of Dap<sup>AMCA</sup>-labeled melittin (MEL<sup>FL</sup>) in cancer cells demonstrated high membrane-penetrating activity, with strong nuclear and nucleolar localization ability. These findings provide implications for novel anticancer therapies based on Trp-substituted designs and nuclear/nucleolar targeted therapy.https://www.mdpi.com/2072-6651/14/7/428melittinTrpDapAMCAfluorescencealpha helix |
| spellingShingle | Yonghui Lv Xu Chen Zhidong Chen Zhanjun Shang Yongxiao Li Wanting Xu Yuan Mo Xinpei Wang Daiyun Xu Shengbin Li Zhe Wang Meiying Wu Junqing Wang Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells Toxins melittin Trp Dap AMCA fluorescence alpha helix |
| title | Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells |
| title_full | Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells |
| title_fullStr | Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells |
| title_full_unstemmed | Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells |
| title_short | Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells |
| title_sort | melittin tryptophan substitution with a fluorescent amino acid reveals the structural basis of selective antitumor effect and subcellular localization in tumor cells |
| topic | melittin Trp Dap AMCA fluorescence alpha helix |
| url | https://www.mdpi.com/2072-6651/14/7/428 |
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