Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells.
Melittin Tryptophan Substitution with a Fluorescent Amino Acid Reveals the Structural Basis of Selective Antitumor Effect and Subcellular Localization in Tumor Cells.
复制标题
用荧光氨基酸取代蜂毒肽色氨酸揭示了肿瘤细胞中选择性抗肿瘤作用和亚细胞定位的结构基础
作者：
蜂毒素是一种膜活性多肽，具有很强的抗癌活性。尽管经过了几十年的研究，但单一色氨酸在蜂毒素的抗癌活性和选择性中的作用仍然知之甚少。在这里，我们提出了一种基于Trp19被非规范荧光氨基酸(DapAMCA)取代的治疗方案。圆二色谱和分子动力学模拟表明，在蜂毒素中引入DapAMCA残基稳定了多肽的螺旋结构。体外溶血和抗癌活性分析表明，在蜂毒素中引入DapAMCA残基改变了其与细胞膜的作用方式，从而降低了溶血毒性，提高了选择性指数(SI)，与蜂毒素相比提高了5倍。体外荧光成像显示DapAMCA标记的蜂毒素(MELFL)在癌细胞中具有很高的穿膜活性，具有很强的核仁定位能力。这些发现为基于色氨酸取代设计和核/核仁靶向治疗的新型抗癌治疗提供了启示。
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 (DapAMCA). The introduction of DapAMCA 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 DapAMCA 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 DapAMCA-labeled melittin (MELFL) 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.
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DOI：
10.1073/pnas.1307010110
发表时间：
2013-08-27
影响因子：
11.1
作者：
Lee, Ming-Tao;Sun, Tzu-Lin;Huang, Huey W.
通讯作者：
Huang, Huey W.
影响因子：
4.4
作者：
ESSMANN, U;PERERA, L;PEDERSEN, LG
通讯作者：
PEDERSEN, LG
影响因子：
2.9
作者：
GREENFIE.N;FASMAN, GD
通讯作者：
FASMAN, GD
影响因子：
3.4
作者：
Bernèche, S;Nina, M;Roux, B
通讯作者：
Roux, B
影响因子：
2.4
作者：
Ladokhin, Alexey S.;Fernandez-Vidal, Monica;White, Stephen H.
通讯作者：
White, Stephen H.