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

线虫 PIE-1 ZF2 的结构特征

• 批准号: 6406371
• 负责人: SARAH L MICHEL
• 金额: $ 3.33万
• 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/6406371
• 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是一种在秀丽隐杆线虫早期胚胎细胞中发现的蛋白质，与细胞与体细胞的分化有关。 PIE-1包含两个推定的锌结合结构域ZF1和ZF2，后者已显示出在生殖细胞中定位的PIE-1并与P颗粒结合。该研究建议的长期目标是对PIE-1在胚胎开发中的作用有基本的了解。了解负责细胞分化和发育的生化机制对于理解包括癌症和发育障碍在内的人类疾病至关重要。在该团契的支持下，将在合成和重组中准备与PIE-1的ZF2相对应的肽。将使用钴作为光谱探针评估肽的螯合锌的能力，并确定钴和锌的结合常数。使用多维NMR光谱法（例如Coesy，Noesy，Toscy），将解决三维溶液结构。这些特征以及与ZF1结构的比较将使结构/功能关系得以评估。