Identification and Evaluation of the Minimum Unit of a KALA Peptide Required for Gene Delivery and Immune Activation
Identification and Evaluation of the Minimum Unit of a KALA Peptide Required for Gene Delivery and Immune Activation
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基因传递和免疫激活所需的 KALA 肽最小单位的鉴定和评估
DOI：
10.1016/j.xphs.2017.05.014
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发表时间：
2017
影响因子：
3.8
通讯作者：
Akita Hidetaka
中科院分区：
文献类型：
--
作者：
Miura Naoya;Tange Kota;Nakai Yuta;Yoshioka Hiroki;Harashima Hideyoshi;Akita Hidetaka
KALA 肽 (WEAKLAKALAKALAKHLAKALAKALKA) 是一种两亲性肽，在生理 pH 值下形成 α 螺旋结构。我们之前报道过，当用 KALA 肽修饰包裹质粒 DNA 的脂质体膜时，可以促进骨髓源性树突状细胞 (BMDC) 中的转基因表达和免疫激活。然而，KALA 肽的最小单位及其二级结构对这些活性的重要性目前尚不完全清楚。我们在此报告了活性所需的 KALA 肽（短 KALA）最小单位的鉴定，通过逐步去除“K-A-L-A”单位来确定。我们通过修饰包裹质粒DNA的多功能包膜型纳米器件来评估4种短KALA的活性。在测试的肽中，短 KALA3 (WEAKLAKALAKALA) 是最短的 KALA 肽，可以形成 α 螺旋结构，并在 BMDC 中引发转基因表达和免疫激活。此外，短 KALA3 作为细胞摄取诱导剂的功能被保留，而在更短的 KALA 版本（短 KALA4）中，摄取完全丧失，因为转基因表达和免疫激活都完全丧失。这些集体数据表明，KALA 肽必须形成 α-螺旋结构才能诱导 BMDC 的细胞摄取。
The KALA peptide (WEAKLAKALAKALAKHLAKALAKALKA) is an amphiphilic peptide that forms an α-helical structure at physiological pH. We previously reported that, when a plasmid DNA-encapsulating liposomal membrane is modified with the KALA peptide, transgene expression and immune activation are facilitated in bone marrow-derived dendritic cells (BMDCs). However, the minimum unit of the KALA peptide and the importance of its secondary structure for these activities are not completely known at this time. We herein report on the identification of the minimum unit of the KALA peptide (short-KALA) required for activity, as determined by the stepwise removal of “K-A-L-A” units. We evaluated the activities of 4 types of short-KALAs by modifying plasmid DNA-encapsulating multi-functional envelop-type nano devices. Among the peptides tested, a short-KALA3 (WEAKLAKALAKALA) was the shortest KALA peptide that could form an α-helical structure, as well as to elicit transgene expression and immune activation in BMDCs. Furthermore, the function of the short-KALA3 as an inducer of cellular uptake was retained, while uptake was completely lost in more shortened versions of KALA (short KALA4), in that transgene expression and immunological activation were both completely lost. These collective data show that the KALA peptide must form an α-helical structure to induce cellular uptake in BMDCs.