EAGER: Engineering bacterial lectins for glycomics analysis

糖基化是将聚糖附着到蛋白质、脂质或其他有机分子上的过程。蛋白质和脂质上的聚糖可以在许多重要的生物过程中作为标记或标识，如细胞生长、细胞-细胞相互作用和细胞迁移。不寻常的糖基化与包括癌症在内的许多不同类型的疾病有关。缺乏简单有效的检测糖基化的工具，这对这一领域的进展造成了重大障碍。PI提出了一项研究，以开发新的工具和方法，有助于低成本和简单易用的糖基化检测。提出了两个具体目标：目标1:PI提出开发和验证一种新的糖蛋白定量方法，该方法可以纳入常用的蛋白质分离技术。如果成功，本研究将扩展一维和二维电泳分离技术的应用。它还可以使这些工具更可靠地用于糖科学和有用的工具。目标2：作为一个例子，PI提出证明所开发的方法可用于实时无标记细菌表面糖基化分析和研究细菌糖基化对环境条件的响应动力学。该提案还计划将工程凝集素分子纳入常见的蛋白质分离和分析格式，如SDS凝胶电泳。如果成功，这将改变聚糖的检测方式，消除对抗体和其他昂贵试剂的需要。提出的研究可能会促进急需的工具的发展，以简化糖科学研究中常用的方法。该研究的成功有可能使糖糖分析变得不那么费力和耗时，减少劳动和材料密集。因此，糖糖分析可能不再仅由少数专业实验室进行，而可能更容易由普通实验室进行。因此，该研究将为糖科学研究人员带来低成本的工具，对该领域产生重大影响。

1452290ChenGeorgia TechGlycosylation is the process that attaches glycans to proteins, lipids, or other organic molecules. Glycans on proteins and lipids may serve as markers or identifiers in many vital biological processes such as cell growth, cell-cell interactions,and cell mig. Unusual glycosylation has been implicated in many different types of diseases including cancer. The lack of simple and effective tools to detect glycolsylation creates a significant obstacle to the advances in this area. The PI proposes a study to develop new tools and methods useful to assist in the detection of glycolsylation that are low cost and simple to use. Two specific aims are proposed: Aim 1: the PI proposes to develop and validate a novel glycoprotein quantification method, which can be incorporated into commonly used protein separation techniques. If successful the work proposed in could extend the use of 1-D and 2-D electrophoresis separation techniques. It could also make these tools more reliably employed and useful tool for glycoscience. Aim 2: As an example the PI proposes to demonstrate that the developed methods could be used for a real-time label-free bacterial surface glycome profiling and for study of dynamics of bacterial glycosylation in response to environmental conditions. The proposal also plans to incorporate engineered lectin molecules into the common protein separation and analysis format, such as SDS gel electrophoresis. If successful, this will change how glycans are detected by eliminating the need for antibody and other expensive reagents commonly in the processes. The proposed research may advance the development of much needed tools to simplify methods commonly used in glycoscience research. The success of the research has the potential to make glycomic analysis less arduous and less time consuming, less labor and material intensive. As a result, glycomic analysis may no longer be conducted by only a small number of specialized laboratories and may become much more accessible to a common laboratory. Thus the research will have significant impact on the field by bringing low cost tools to glycoscience researchers.