Isolation of novel cell-penetrating peptides from a random peptide library using in vitro virus and their modifications
Isolation of novel cell-penetrating peptides from a random peptide library using in vitro virus and their modifications
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DOI：
10.3892/ijmm_00000311
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发表时间：
2010-01-01
影响因子：
5.4
通讯作者：
Fukamizu, Akiyoshi
中科院分区：
文献类型：
--
作者：
Kamide, Kaeko;Nakakubo, Hiroshi;Fukamizu, Akiyoshi
已经报道了许多细胞穿透肽（CPP），但是它们的转导效率太低而不能用作用于治疗目的的细胞内载体。我们使用体外病毒（IVV）文库进行了全面的搜索以发现新的CPP，所述文库呈现由15个氨基酸组成的随机肽（文库的多样性>10（12））。我们发现了9种新的CPP，其胞内转运效率高于达特肽（YGRKKKRRQRRR）。有趣的是，新CPP之一，No.14（KLWMRWYSPTTRRYG），在CHO细胞中显示出相对于达特肽的易位活性的显著改善（在50 μ M中>10倍的效率）。由于用Arg取代Lys（1）（14-1）提高了No.14的胞内转运效率，我们在14-1的基础上进行了丙氨酸扫描，以确定胞内转运的重要氨基酸。Ala取代分析表明，Arg和Trp残基对穿膜活性都有重要作用，其贡献大小顺序为Trp（3）
A number of cell-penetrating peptides (CPPs) have been reported, but their transduction efficiencies are too low to be used as intracellular carriers for therapeutic purposes. We conducted a comprehensive search to find novel CPPs using an in vitro virus (IVV) library, which presented random peptides consisting of 15 amino acids (diversity of the library was >10(12)). We found 9 kinds of novel CPPs with an intracellular translocation efficiency higher than that of the TAT peptide (YGRKKKRRQRRR). Interestingly, one of the novel CPPs, No. 14 (KLWMRWYSPTTRRYG), showed a dramatic improvement in translocation activity relative to the TAT peptide in CHO cells (>10-fold efficiency in 50 mu M). As the intracellular translocation efficiency of No. 14 was increased by substitution Arg for Lys(1) (14-1), we carried out alanine scanning on the basis of 14-1 to determine important amino acids for the intracellular translocation. The Ala substitution analysis showed that both Arg and Trp residues were important for the cell-penetrating activity and that their contribution was in the order Trp(3)