Development of a novel catalytic array

描述（由申请人提供）：将生成约100种被称为去泛素化酶（DUBs）的蛋白质的生物信息学工具，这些蛋白质在细胞存活、增殖和分化中起着重要作用。lifessensor专有的SUMO表达增强技术允许相对容易地表达和纯化dub，并且这些蛋白质的高通量微阵列将提供快速和经济有效的筛选。因此，建议开发DUB微阵列用于基础研究，生物标志物发现，诊断/预后和治疗。DUB微阵列将在生物标志物发现和诊断开发中具有明显的实用性。dub的表达与癌症、糖尿病和帕金森病等疾病有关。疾病状态下蛋白质的过表达或过低表达往往会引起针对这些蛋白质的自身抗体产生的升高或降低。因此，自身抗体谱分析可以帮助识别疾病的类型和阶段。当一种生物标志物被确定后，可以进一步研究其在疾病诊断和预后中的作用。这种特殊的生物标记物也可能成为治疗靶点。DUB微阵列不仅可以对样品进行高通量筛选，还可以评估相同条件下样品中DUB的表达水平，以关联它们在不同情况下对彼此表达水平的影响。在第一个目标中，将10（10）个dub固定在各种固体载体上，以确定在酶活性保留方面的最佳载体。接下来，将保留的dub的催化功能与它们在标准分析系统中的功能进行比较。最后，50（50）个来自普通人群的血清样本将被分析为针对dub的自身抗体表达的变化。在第二阶段，将构建由所有已知的泛素和相关泛素样蛋白DUBS组成的阵列。

DESCRIPTION (provided by applicant): Bioinformatics tools will be generated for a class of ~ 100 proteins known as de- ubiquitinating enzymes (DUBs), which play an important role in cell survival, proliferation, and differentiation. LifeSensor's proprietary SUMO expression enhancement technology permits expression and purification of DUBs with relative ease, and a high throughput microarray of these proteins will afford rapid and cost effective screening. Thus, it is proposed to develop a DUB microarray for the purpose of basic research, biomarker discovery, diagnostics/prognostics, and therapeutics. The DUB microarray will have obvious utility in biomarker discovery and diagnostics development. Expression of DUBs has been associated with diseases such as cancer, diabetes, and Parkinsonism. Over- or under-expression of proteins in disease states tend to elicit elevated or diminished auto-antibody production against those proteins. Thus, auto-antibody profiling can assist in identifying the type and stage of a disease. When a biomarker is identified, it can be further studied for its role in the diagnosis and prognosis of the disease. The particular biomarker could also be a therapeutic target. A DUB microarry not only allows one to perform high throughput screening of samples, but also to evaluate expression levels of DUBs in samples under identical conditions to correlate their effect on each other's expression levels under different circumstances. In the first aim, 10 (ten) DUBs will be immobilized on a variety of solid supports to determine the optimum support in terms of retention of enzyme activity. Next, the catalytic function of the retained DUBs will be compared with their function in a standard assay system. Finally, 50 (fifty) serum samples from the general population will be profiled for variation in the expression of autoantibodies against the DUBs. In Phase II, arrays will be built consisting of all known DUBS for ubiquitin and relevant ubiquitin-like proteins.