Structural Characterization of ZF2 of PIE-1 in C.elegans

线虫 PIE-1 ZF2 的结构特征

• 批准号: 6607181
• 负责人: SARAH L MICHEL
• 金额: $ 4.64万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-02 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6607181
• 关键词: Caenorhabditis elegans; binding sites; chelating agents; cobalt; computer simulation; embryo /fetus protein; intermolecular interaction; ionic bond; metal complex; metalloproteins; model design /development; molecular dynamics; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; physical model; protein localization; protein sequence; protein structure function; recombinant proteins; structural biology; surface property; zinc

PIE-1, a protein found in early embryonic cells of C. elegans, is involved in the differentiation of germ cells from somatic cells. PIE-1 contains two putative zinc binding domains, ZF1 and ZF2, the latter of which has been shown to localize PIE-1 in germ cells and bind to P granules. The long term goal of this research proposal is to gain a fundamental understanding of the role of ZF2 of PIE-1 in embryonic development. Understanding the biochemical mechanisms responsible for cell differentiation and development is crucial for understanding human disorders including cancer and developmental disorders. With the support of this fellowship, a peptide that corresponds to ZF2 of PIE-1 will be prepared, both synthetically and recombinantly. The peptide's ability to chelate zinc will be assessed using cobalt as a spectroscopic probe and binding constants for both cobalt and zinc will be determined. A three dimensional solution structure will be solved, using multi-- dimensional NMR spectroscopy (e.g. COESY, NOESY, TOSCY). These characterizations coupled with comparisons to the ZF1 structure will enable structure/function relationships to be assessed.

PIE-1 是一种在线虫早期胚胎细胞中发现的蛋白质，参与生殖细胞从体细胞的分化。 PIE-1 包含两个假定的锌结合结构域，ZF1 和 ZF2，后者已被证明将 PIE-1 定位在生殖细胞中并与 P 颗粒结合。本研究计划的长期目标是从根本上了解 PIE-1 的 ZF2 在胚胎发育中的作用。了解负责细胞分化和发育的生化机制对于了解包括癌症和发育障碍在内的人类疾病至关重要。在该奖学金的支持下，将通过合成和重组制备与PIE-1的ZF2相对应的肽。将使用钴作为光谱探针评估肽螯合锌的能力，并确定钴和锌的结合常数。将使用多维核磁共振波谱（例如 COESY、NOESY、TOSCY）求解三维溶液结构。这些表征加上与 ZF1 结构的比较将能够评估结构/功能关系。