Ganglioside interactome toolkit

神经节苷脂，唾液酸化鞘糖脂，存在于所有脊椎动物细胞和组织中， 在影响人类疾病（包括糖尿病、癌症、神经系统疾病）的多种分子信号通路中发挥作用， 变性蛋白质病、智力残疾和许多其他疾病。主要的神经节苷脂结构是 定义明确、有限且在脊椎动物物种间共享。它们的聚糖独立调节细胞信号 一种蛋白质载体。大多数神经节苷脂存在于细胞表面，其神经酰胺脂质嵌入细胞膜中。 质膜和它们的聚糖向外延伸。它们以两种方式调节细胞生理学，顺式和 译作为顺式调节剂，它们通过聚糖结合到相同细胞中的跨膜蛋白而横向缔合。 细胞膜调节其功能。作为反式识别分子，它们与蛋白质结合， 细胞外环境或并置细胞上，介导细胞-细胞相互作用。顺式和反式相互作用都是 对神经节苷脂聚糖结构具有特异性，对人类健康至关重要。 大多数神经节苷脂的功能和神经节苷脂-蛋白质相互作用仍然知之甚少，因为缺乏 广泛使用、适应性强和经过验证的工具以及优化的使用方法。这是我们的目标 利用化学生物学技术解决这一需要的项目，以合成一套确定的主要 神经节苷脂携带最小破坏性的双功能光反应和炔（点击化学）标签。 将生成优化和验证的方案，以将双功能标记的神经节苷脂递送到外部细胞。 细胞质膜的小叶，大多数神经节苷脂居住和发挥功能。我们的目标是 合成神经节苷脂探针工具包，验证探针的适当细胞递送，验证它们用于 鉴定聚糖特异性神经节苷脂结合蛋白，并将试剂和方案转移到其他 生物医学研究实验室在这个项目的期限内。可交付成果、神经节苷脂探针和 经验证的方法，将广泛分发给生物医学研究人员，以发现身份， 与多种人类细胞相关的神经节苷脂结合蛋白的特异性、分布和功能， 组织和疾病。

Gangliosides, sialylated glycosphingolipids found on all vertebrate cells and tissues, play well-established roles in diverse molecular signaling pathways that impact human diseases including diabetes, cancer, neuro- degenerative proteinopathies, intellectual disability, and many others. The major ganglioside structures are well-defined, finite, and shared across vertebrate species. Their glycans regulate cell signaling independent of a protein carrier. Most gangliosides reside on the cell surface with their ceramide lipids embedded in the plasma membrane and their glycans extending outward. They regulate cell physiology in two modes, cis and trans. As cis regulators, they associate laterally via glycan binding to transmembrane proteins in the same cell membrane to regulate their function. As trans recognition molecules they engage proteins in the extracellular milieu or on apposing cells, mediating cell-cell interactions. Both cis and trans interactions are specific for ganglioside glycan structures and essential for human health. Most ganglioside functions and ganglioside-protein interactions remain poorly understood due to lack of broadly accessible, adaptable and validated tools and optimized methods for their use. It is the goal of this project to address this need using chemical biology technologies to synthesize a defined set of major gangliosides carrying minimally disruptive bifunctional photoreactive and alkyne (click chemistry) tags. Optimized and validated protocols will be generated to deliver bifunctionally tagged gangliosides to the outer leaflet of the plasma membrane of cells, where most gangliosides reside and function. Our goal is to synthesize the ganglioside probe toolkit, validate appropriate cell delivery of the probes, validate their use to identify glycan-specific ganglioside binding proteins, and transfer the reagents and protocols to other biomedical research laboratories within the term of this project. The deliverables, ganglioside probes and validated methods for their use, will be distributed broadly to biomedical researchers to discover the identities, specificities, distributions and functions of ganglioside binding proteins relevant to a variety of human cells, tissues, and diseases.