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

PIE-1是一种在线虫早期胚胎细胞中发现的蛋白质，参与了生殖细胞和体细胞的分化。PIE-1含有两个可能的锌结合结构域，ZF1和ZF2，后者被证明定位于生殖细胞并与P颗粒结合。这项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理解包括癌症和发育障碍在内的人类疾病至关重要。在该基金的支持下，将合成和重组与PIE-1的ZF2相对应的多肽。将使用钴作为光谱探针来评估多肽对锌的螯合能力，并将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)来求解三维溶液结构。这些特征加上与ZF1结构的比较，将使结构/功能关系得以评估。

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.