Characterization of p53-mediated cell death in neurons

产品说明： 转录激活因子和抑制因子p53调节神经元活力 在遗传毒性或兴奋性毒性损伤后， 我们对此知之甚少。为了阐明p53依赖的细胞死亡， 机制，我们将使用新开发的技术，涉及傅立叶变换 电离回旋共振质谱和微分稳定 半胱氨酸残基的同位素标记以表征蛋白质组（整个 在感兴趣的细胞或组织中的一组表达蛋白质 神经元在对照条件下或用谷氨酸或 DNA损伤剂这种方法的速度和灵敏度超过了 目前可用的2D-PAGE方法的几个数量级。的 数以千计的蛋白质的身份和表达水平，包括低 丰富和以前未知的物种，将迅速获得复杂的 从培养的神经元中提取的蛋白质混合物。腺病毒 p53、p53特异性抑制剂和p53敲除小鼠的过表达将被 用于操纵p53水平并寻找蛋白质表达的变化。

DESCRIPTION: The transcriptional activator and repressor, p53, modulates neuronal viability after genotoxic or excitotoxic injury, but the mechanisms by which this occurs are poorly understood. In order to elucidate p53-dependent cell death mechanisms, we will use newly developed technology involving Fourier transform ionization cyclotron resonance mass spectrometry and differential stable isotope labeling of cysteine residues to characterize the proteome (the entire set of expressed proteins in a cell or tissue of interest) of cultured cortical neurons under control conditions or after treatment with glutamate or DNA-damaging agents. The speed and sensitivity of this method exceeds that of currently available 2D-PAGE methods by several orders of magnitude. The identities and expression levels of thousands of proteins, including low abundance and previously unknown species, will be rapidly obtained from complex protein mixtures extracted from cultured neurons. Adenovirus-mediated over-expression of p53, p53-specific inhibitors and p53 knockout mice will be used to manipulate p53 levels and look for changes in protein expression.