RUI: Regulation and Function of Novel CaM-Related Proteins in Arabidopsis

钙调素（CaM）蛋白的功能是将化学信号（细胞溶质钙的增加）转化为生化信号。 CaM蛋白识别靶蛋白并与靶蛋白功能性相互作用的能力在该信号转导过程中是必不可少的。 在许多生物体中，除了植物外，存在一个或两个CaM型蛋白。 在模式生物拟南芥中，有七种已知的CaM蛋白和五种额外的CaM相关蛋白。 对拟南芥基因组的进一步分析表明，至少还有13种潜在的钙调素相关蛋白。 在这个项目中，三个钙调素相关蛋白之间的生物化学和功能差异将得到解决。 这些蛋白质都具有保守的半胱氨酸残基，这被认为是形成二硫键。 本PI将研究半胱氨酸残基和潜在二硫键形成在钙结合和与疏水性树脂结合中的作用。 如果二硫键的形成似乎影响蛋白质结合钙和与疏水表面相互作用的能力，则半胱氨酸残基将单独突变。 然后表达和纯化突变的蛋白质，以测试消除特定半胱氨酸残基对这些蛋白质的生化活性的影响

Calmodulin (CaM) proteins function to transduce a chemical signal (increase in cytosolic calcium) into a biochemical signal. The ability of the CaM protein to recognize and functionally interact with a target protein is essential in this signal transduction process. In many organisms, with the exception of plants, there exist one or two CaM-type proteins. In the model organism Arabidopsis thaliana there are seven known CaM proteins and five additional CaM-related proteins. Further analysis of the Arabidopsis genome suggests that there are at least an additional 13 potential CaM-related proteins. In this project, the biochemical and functional difference between three CaM-related proteins will be addressed. These proteins all possess conserved cysteine residues, which are proposed to form a disulfide bond. This PI will investigate the role of the cysteine residues and potential disulfide bond formation in calcium binding and in binding to a hydrophobic resin. If disulfide bond formation appears to influence the ability of the proteins to bind calcium and to interact with hydrophobic surfaces, the cysteine residues will be individually mutated. The mutated protein will then be expressed and purified to test the effect of elimination of specific cysteine residues on the biochemical activity of these proteins