Direct protein delivery into intact plant cells using polyhistidine peptides
Direct protein delivery into intact plant cells using polyhistidine peptides
复制标题
DOI：
10.1093/bbb/zbab055
复制
发表时间：
2021-03-31
影响因子：
1.6
通讯作者：
Iwasaki, Takashi
中科院分区：
文献类型：
--
作者：
Tanaka, Yoshino;Nanasato, Yoshihiko;Iwasaki, Takashi
多组氨酸肽（Polyhistidine peptides，PHPs）是一类仅含组氨酸残基（> His 8）的多肽，是植物细胞中有效的细胞穿透肽。利用PHP融合蛋白，我们的目的是将蛋白质传递到培养的烟草，水稻和柳杉植物细胞。将培养的细胞与在一种多肽中结合麦芽糖结合蛋白（MBP）、红色荧光蛋白（RFP）和各种PHP的融合蛋白（MBP-RFP-His 8-His 20）共培养，显示融合蛋白在所有植物细胞系中的细胞摄取。最大细胞内荧光显示在MBP-RFP-His 20中。此外，腺苷酸环化酶（CyaA），一种由胞质钙调蛋白激活的环磷酸腺苷（cAMP）合酶，被用作活细胞中蛋白质递送的报告基因。将MBP、RFP、CyaA和His 20（MBP-RFP-CyaA-His 20）组合的融合蛋白递送到植物细胞中，并在所有细胞系中增加细胞内荧光和cAMP产生。本研究表明，PHPs是各种培养植物细胞的胞内空间的蛋白质的有效载体。
Polyhistidine peptides (PHPs), sequences comprising only histidine residues (>His8), are effective cell-penetrating peptides for plant cells. Using PHP-fusion proteins, we aimed to deliver proteins into cultured plant cells from Nicotiana tabacum, Oryza sativa, and Cryptomeria japonica. Co-cultivation of cultured cells with fusion proteins combining maltose-binding protein (MBP), red fluorescent protein (RFP), and various PHPs (MBP-RFP-His8-His20) in one polypeptide showed the cellular uptake of fusion proteins in all plant cell lines. Maximum intracellular fluorescence was shown in MBP-RFP-His20. Further, adenylate cyclase (CyaA), a synthase of cyclic adenosine monophosphate (cAMP) activated by cytosolic calmodulin, was used as a reporter for protein delivery in living cells. A fusion protein combining MBP, RFP, CyaA, and His20 (MBP-RFP-CyaA-His20) was delivered into plant cells and increased intracellular fluorescence and cAMP production in all cell lines. The present study demonstrates that PHPs are effective carriers of proteins into the intracellular space of various cultured plant cells.