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Design and intracellular delivery of peptides for transcription regulation

用于转录调控的肽的设计和细胞内递送

基本信息

批准号：
12557200
负责人：
FUTAKI Shiroh
金额：
$ 8.58万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557200/
关键词：
signal transduction synthetic peptide HIV translocation peptide engineering transcription regulation transcription foctor arginine 細胞膜透過ペプチドタンパク質細胞内導入 HIV-1 Tat HIV-1 Rev 薬物送達核移行

项目摘要

A basic peptide derived from the human immunodeficiency virus (HIV)-1 Tat has been reported to have the ability to translocate through the cell membranes and to bring exogenous proteins into the cells. We have demonstrated that these features were observable among many arginine-rich peptides including those having a branched chain structure. Based on these findings, the presence of a ubiquitous internalization mechanism for the arginine-rich peptides has been suggested. We have also demonstrated that these features are also applicable to the peptides having branched-chain structures. Peptides that have arginine residues on four branched-chains (R_n)_4 [n (number of arginine residues) = 0〜6] were prepared. Fluorescence microscopic observation revealed that the (R_2)_4 peptide showed the most efficient translocation. Dependence on the number of arginine residues for the translocation efficiency and cellular localization was also observed for the branched-chain peptides as was seen in the linear peptides. We have shown that a non-covalent protein assembly of Rnase S bearing arginine-rich segment was successfully introduced into cells to exhibit an anti-HIV activity. Peptides corresponding to the phosphorylation and ubiquitilation sites of IκB, which is involved in the activation of transcription factor NF-κB, were prepared. Introduction of these peptides into cells by conjugation with the membrane-permeable arginine peptide resulted in the inhibition of NF-κB activation. However, significance of the difference in the extent of inhibition was observed. We have also shown that the transcription by transcription factor Sp1 was inhibited by the peptide derived from DNA recognition segment of Sp1.

据报道，源自人类免疫缺陷病毒（HIV）-1达特的碱性肽具有穿过细胞膜并将外源蛋白带入细胞的能力。我们已经证明，这些功能是可观察到的许多富含精氨酸的肽，包括那些具有支链结构。基于这些发现，已经提出了富含精氨酸的肽的普遍存在的内化机制的存在。我们还证明了这些特征也适用于具有支链结构的肽。制备了四个支链（Rn）_4 [n（精氨酸残基数）= 0 ~ 16]的精氨酸残基肽。荧光显微镜观察表明，（R_2）_4肽的转运效率最高。对于支链肽，也观察到对于易位效率和细胞定位的精氨酸残基数目的依赖性，如在线性肽中所见。我们已经证明，RNA酶S的非共价蛋白组装体携带富含精氨酸的片段被成功地引入细胞中，以表现出抗HIV活性。制备了与IκB的磷酸化和泛素化位点相对应的肽，IκB参与转录因子NF-κB的活化。通过与膜渗透性精氨酸肽缀合将这些肽引入细胞导致NF-κB活化的抑制。然而，观察到抑制程度的显著差异。我们还发现转录因子Sp1的转录被Sp1的DNA识别片段衍生的肽抑制。