HYDROLYTIC CLEAVAGE OF DNA BY METAL-PEPTIDE INTERCALATOR

通过金属-肽嵌入剂对 DNA 进行水解切割

• 批准号: 2668445
• 负责人: ROBERT P HOUSER
• 金额: $ 2.62万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2668445
• 关键词: DNA; calcium ion; chemical synthesis; cobalt; copper; divalent cations; hydrolysis; lead; ligands; magnesium; magnesium ion; metal complex; mutagens; nucleic acid metabolism; oligonucleotides; peptides; phenanthrene; quinones; zinc

Construction of artificial nucleases is an attractive goal, having implications in molecular biology, genetic engineering and clinical medicine. The proposed research aims to answer important questions regarding the hydrolytic cleavage of DNA by investigating new molecules designed to hydrolytically cleave the DNA double-helix. In particular, a series of rhodium metallointercalators with covalently appended synthetic peptides which contain metal binding sites will be prepared. The proposed complexes are based upon recently prepared metal-peptide conjugates, Rh(phi)2L (phi = phenanthrenequinone diimine; L = bidentate ligands with a covalently appended peptide). The appended peptide will be systematically varied by introducing amino acid residues such as histidine, cysteine, or glutamate into the sequence, which will act as ligands, creating a coordination site where various divalent metal ions can bind, and potentially interact with and cleave the sugarphosphodiester backbone of DNA. Using the rhodium-peptide chimeras in the presence of divalent metal ions, the DNA cleavage reaction will be investigated, and the conditions for hydrolytic scission of the phosphate diester bond will be optimized. The new complexes will be fully characterized using multiple spectroscopic and physical methods. Analysis of the information gleaned from the characterization of the rhodium- peptide-metal species will allow the correlation of factors such as peptide secondary structure, metal ion type and affinity, coordinating ligands, and DNA affinity with hydrolysis. Through the study of the reactions of these complexes with DNA, important insights into DNA hydrolysis will be gained. As a result of these studies, significant progress toward the creation of artificial nucleases could be gained.

构建人工核酸酶是一个有吸引力的目标，具有 分子生物学、基因工程和临床的意义 医药。这项拟议的研究旨在回答一些重要问题 关于DNA的水解性切割的新研究 被设计用来水解性切割DNA双螺旋的分子。 特别是一系列具有共价结构的金属铑嵌入剂。 附加的含有金属结合位点的合成肽将是 准备好了。建议的复合体是基于最近准备的 金属-多肽结合物，Rh(Phi)2L(phi=菲醌 二亚胺；L=带有共价附加多肽的双齿配体)。 附加的多肽将通过引入 氨基酸残基，如组氨酸、半胱氨酸或谷氨酸 序列，它将作为配体，创建一个配位位点 不同的二价金属离子可以结合并潜在地相互作用 结合并切割DNA的糖磷酸二酯骨架。vbl.使用 在二价金属离子存在的情况下形成的Rg-多肽嵌合体， 将对DNA切割反应进行研究，并在 对于磷酸二酯键的水解性断裂 最优化。新的络合物将被充分表征使用 多种光谱和物理方法。三、分析了 从铑的特征中收集到的信息- 多肽-金属物种将允许相关因素，如 多肽二级结构，金属离子类型和亲和力，配位 配体，以及DNA与水解物的亲和力。通过对这一问题的研究， 这些络合物与DNA的反应，对 DNA的水解度将得到提高。作为这些研究的结果， 在创造人造核酸酶方面取得重大进展可能 被获得了。