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

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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.