Measurement of nitric-oxide-modified proteins by cavity ring-down spectroscopy

说明书(由申请人提供)一氧化氮合酶产品通过与金属蛋白中的血红素中心相互作用，例如鸟氨酸环化酶，以及通过与蛋白质半胱氨酸硫醇的共价反应来发出信号。这些亚硝化反应与健康和疾病中的真核细胞生物学有着广泛的相关性。然而，临床和研究样本所需的灵敏度和特异度往往难以检测到硝基蛋白质。我们正在开发腔衰荡光谱来测量这些蛋白质修饰。我们相信，这项技术将极大地提高灵敏度和特异度，它有可能给该领域带来革命性的变化。在这里，我们建议设计和制造一种具有样品光解界面的改进的腔衰减仪。该仪器将使用先进的光谱技术来测定生物标本中的S-亚硝硫醇。它将被用来研究广泛的疾病和器官系统。它将比以前的技术更强大、更精确，以最小的样品干扰准确地测量PM浓度中未修饰的蛋白质。在第二个目标中，它还将应用于蛋白质组微阵列，以实现快速、高通量的发现。

DESCRIPTION (provided by applicant) Nitric oxide synthase products signal through interactions with heme centers in metalloproteins, such as guanylate cyclase and through covalent reactions with protein cysteine thiols. These nitrosylation reactions have broad relevance to eukaryotic cell biology in health and disease. However, nitrosylated proteins have often proven difficult to detect with the sensitivity and specificity needed for clinical and reseach samples. We are developing cavity ring-down spectroscopy to measure these protein modifications. We believe that this technology will dramatically improve both sensitivity and specificity it has the potential to will revolutionize the field. Here, we propose to design and fabricate an improved cavity ring-down instrument with a sample photolysis interface. This instrument will use advanced optical spectroscopy techniques to measure S- nitrosothiols in biological specimens. It will be used to studying a broad range of diseases and organ systems. It will be more powerful and more precise than previous technologies, measuring NO-modified proteins accurately in pM concentrations with minimal sample disruption. In the second Aim, it will also be applied to proteomic microarrays for rapid, high-throughput discovery.