Synthetic Mitochondria-Targeting Peptides Incorporating α-Aminoisobutyric Acid with a Stable Amphiphilic Helix Conformation in Plant Cells
Synthetic Mitochondria-Targeting Peptides Incorporating α-Aminoisobutyric Acid with a Stable Amphiphilic Helix Conformation in Plant Cells
复制标题
植物细胞中包含具有稳定两亲螺旋构象的 α-氨基异丁酸的合成线粒体靶向肽
DOI：
10.1021/acsbiomaterials.0c01533
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发表时间：
2021
影响因子：
5.8
通讯作者：
and Keiji Numata
中科院分区：
文献类型：
--
作者：
Kayo Terada;Joan Gimenez-Dejoz;Taichi Kurita;Kazusato Oikawa;Hirotaka Uji;Kousuke Tsuchiya;and Keiji Numata
在植物细胞的基因改造中，线粒体是除细胞核和质体外的重要靶点。然而，由于线粒体的体积小，流动性高，基因传递到植物细胞的线粒体尚未通过传统的方法，如粒子轰击建立。为了开发一种在植物细胞中有效的线粒体靶向信号（MTS），我们设计了人工肽（LURL）3及其类似物，其周期性特征为疏水性α-氨基异丁酸（Aib， U）和阳离子精氨酸(R)，考虑到线粒体输入受体Tom20识别的一致基序。圆二色性测量和分子动力学模拟研究表明，（LURL）3在含有1.0 wt %十二烷基硫酸钠的0.1 M磷酸盐缓冲溶液中倾向于形成稳定的α-螺旋结构。在通过粒子轰击进入植物细胞后，（LURL）3在线粒体中表现出高度选择性的积累，而它的类似物（LARL）3除了位于线粒体外，还主要位于液泡中。（LURL）3的高选择性可归因于Aib的掺入，Aib通过增加（LURL）3的疏水性和螺旋度，促进了MTS与Tom20之间的疏水相互作用。本研究提供了一个前瞻性的线粒体靶向系统，使用人工肽的简单设计。
In the genetic modification of plant cells, the mitochondrion is an important target in addition to the nucleus and plastid. However, gene delivery into the mitochondria of plant cells has yet to be established by conventional methods, such as particle bombardment, because of the small size and high mobility of mitochondria. To develop an efficient mitochondria-targeting signal (MTS) that functions in plant cells, we designed the artificial peptide (LURL)3and its analogues, which periodically feature hydrophobic α-aminoisobutyric acid (Aib, U) and cationic arginine (R), considering the consensus motif recognized by the mitochondrial import receptor Tom20. Circular dichroism measurements and molecular dynamics simulation studies revealed that (LURL)3had a propensity to form a stable α-helix in 0.1 M phosphate buffer solution containing 1.0 wt % sodium dodecyl sulfate. After internalization into plant cells via particle bombardment, (LURL)3revealed highly selective accumulation in the mitochondria, whereas its analogue (LARL)3was predominantly located in the vacuoles in addition to mitochondria. The high selectivity of (LURL)3can be attributed to the incorporation of Aib, which promotes the hydrophobic interaction between the MTS and Tom20 by increasing the hydrophobicity and helicity of (LURL)3. The present study provided a prospective mitochondrial targeting system using the simple design of artificial peptides.