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: 2021425
• 负责人: ROBERT P HOUSER
• 金额: $ 2.43万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-31 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2021425
• 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.

人工核酸酶的构建是一个有吸引力的目标， 分子生物学、基因工程和临床 药这项研究旨在回答一些重要的问题 关于DNA的水解裂解，通过研究新的 设计用来水解切割DNA双螺旋的分子。 特别地，一系列的铑金属插层剂与共价键 附加的含有金属结合位点的合成肽将 制备所提出的配合物是基于最近制备的 金属-肽缀合物，Rh（phi）2L（phi =菲醌 二亚胺; L =具有共价连接的肽的双齿配体）。 附加的肽将通过引入 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，创造一个协调位点 其中各种二价金属离子可以结合并潜在地相互作用 并切割DNA的糖磷酸二酯骨架。使用 在二价金属离子存在下的铑-肽嵌合体， 将研究DNA切割反应，并且条件 对于磷酸二酯键的水解断裂， 优化新的复合物将充分表征使用 多种光谱和物理方法。分析 从铑的表征中收集的信息- 肽-金属物质将允许以下因素的相关性， 肽二级结构，金属离子类型和亲和力，配位 配体和DNA与水解的亲和力。通过研究 这些复合物与DNA的反应，重要的见解 将获得DNA水解。作为这些研究的结果， 在创造人工核酸酶方面的重大进展可以 获得。