Proteomic Approach to G-Quartets and Cell Aging

G-四重奏和细胞衰老的蛋白质组学方法

• 批准号: 6576620
• 负责人: Linda B. Mc GOWN
• 金额: $ 7.7万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6576620
• 关键词: binding proteins; biotechnology; capillary electrophoresis; cell age; cell senescence; guanine; mass spectrometry; matrix assisted laser desorption ionization; nucleic acid sequence; nucleic acid structure; oligonucleotides; proteomics; telomere; tissue /cell culture

The goal of this R03 project is to develop tools for identification of celt proteins that bind to G-quartet structures formed by telomeric repeat sequences of human chromosomes. The proposed research takes two "directed" proteomic approaches to capture of G-quartet binding proteins from cell lysates. In the first approach, oligonucleotides with telomeric repeat sequences will be immobilized at the surfaces of capillaries for affinity capillary electrophoresis. Captured proteins will then be analyzed by matrix assisted laser desorption/ionization (MALDI) mass spectrometry. In the second approach, the oligonucleotides will be attached to substrate surfaces for surface enhanced laser desorption/ionization (SELDI) mass spectrometric analysis. The long-term goal of the proposed research is to study the role of G-quartet DNA structures in cellular processes related to aging. Telomeres are nucleoprotein complexes that cap the ends of eukaryotic chromosomes, affording protection from degradation by nucleases or fusion with the ends of other chromosomes. The G-rich strands of telomeric DNA form highly stable, intramolecular G-quartet structures in vitro. Although in vivo formation of G-quartet structures by telomeric DNA has yet to be confirmed, there is compelling evidence that G-quartets may play a role in telomere elongation, genetic recombination, and gene regulation. Proteins that bind to G-quartet structures formed by telomeric DNA sequences in vitro, including human DNA topoisomerase and several helicases, have been associated with genetic instability. The proposed research is an essential first step towards elucidation of the relationship between G-quartet binding proteins and telomeric processes in cells.

这个R03项目的目标是开发工具，用于鉴定与人类染色体端粒重复序列形成的g -四重奏结构结合的细胞蛋白。拟议的研究采用两种“定向”蛋白质组学方法从细胞裂解物中捕获g -四重奏结合蛋白。在第一种方法中，端粒重复序列的寡核苷酸将固定在毛细血管表面进行亲和毛细管电泳。然后，捕获的蛋白质将通过基质辅助激光解吸/电离（MALDI）质谱分析。

项目成果

Detection of G-quartet structure in a DNA aptamer stationary phase using a fluorescent dye.

使用荧光染料检测 DNA 适体固定相中的 G 四联体结构。

• DOI: 10.1366/0003702041389364
• 发表时间: 2004
• 期刊: Applied spectroscopy.
• 作者: Joyce,MichelleViglietta;McGown,LindaB
• 通讯作者: McGown,LindaB