Development of a novel catalytic array

描述（由申请人提供）：将为一类称为去泛素化酶（DUB）的约100种蛋白质生成生物信息学工具，这些蛋白质在细胞存活、增殖和分化中发挥重要作用。LifeSensor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具有成本效益的筛选。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/诊断和治疗的目的。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、糖尿病和帕金森综合征等疾病有关。在疾病状态中蛋白质的过度或不足表达倾向于引起针对那些蛋白质的自身抗体产生的升高或降低。因此，自身抗体分析可以帮助识别疾病的类型和阶段。当一个生物标志物被确定，它可以进一步研究其在疾病的诊断和预后中的作用。特定的生物标志物也可以是治疗靶标。DUB微阵列不仅允许对样品进行高通量筛选，而且还可以在相同条件下评估样品中DUB的表达水平，以将它们在不同情况下对彼此表达水平的影响相关联。在第一个目标中，将10（十）个DUB固定在各种固体支持物上以确定在酶活性保留方面的最佳支持物。接下来，将保留的DUB的催化功能与它们在标准测定系统中的功能进行比较。最后，将对来自一般人群的50（五十）份血清样本进行抗DUB自身抗体表达变化的分析。在第二阶段，将建立由所有已知的泛素和相关泛素样蛋白的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.