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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：
14370720
负责人：
FUTAKI Shiroh
金额：
$ 9.15万
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：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

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. 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. The site of action for these complexes resides in the stages between the viral entry into the cells and reverse transcription, suggesting the possibility of the recognition of viral RNA by these basic peptides in the cells. 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. As for cross-linking formation between delivered molecules and cellular proteins, the feasibility of employment of light-induced cross-linker and aldehyde moieties was assessed. However, since the efficiency of cross-linking formation was not satisfactory, we are now seeking alternative systems to resolve this problem.

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。我们已经证明，一个非共价蛋白组装的RNase S轴承富含精氨酸的片段被成功地引入细胞表现出抗HIV活性。这些复合物的作用位点位于病毒进入细胞和逆转录之间的阶段，表明这些碱性肽在细胞中识别病毒RNA的可能性。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。至于递送的分子和细胞蛋白质之间的交联形成，评估了采用光诱导交联剂和醛部分的可行性。然而，由于交联形成的效率不令人满意，我们现在正在寻找替代系统来解决这个问题。