Ganglioside interactome toolkit

神经节苷脂相互作用组工具包

• 批准号: 9813609
• 负责人: RONALD L SCHNAAR
• 金额: $ 49.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9813609
• 关键词: Address; Alkynes; Azides; Binding; Binding Proteins; Biology; Biomedical Research; Biotin; Carrier Proteins; Cell Communication; Cell Culture Techniques; Cell membrane; Cell physiology; Cell surface; Cells; Ceramides; Charge; Chemicals; Chemistry; Diabetes Mellitus; Diazomethane; Disease; Endocytosis; Gangliosides; Glycosphingolipids; Goals; Health; Human; Hydrogen Bonding; Integral Membrane Protein; Intellectual functioning disability; Intracellular Membranes; Laboratories; Laboratory Research; Link; Lipids; Malignant Neoplasms; Mediating; Methanol; Methods; Minority; Molecular; Myelin Associated Glycoprotein; Nerve Degeneration; Nervous system structure; Neurodegenerative Disorders; Outcome; Play; Polysaccharides; Production; Proteins; Proteome; Proteomics; Protocols documentation; Reagent; Research Personnel; Role; Sialic Acids; Signal Pathway; Signal Transduction; Site; Specificity; Structure; Surface; System; Technology; Tissues; Validation; experimental study; extracellular; fluorophore; functional group; human disease; interest; metabolomics; tool

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.

神经节苷脂是在所有脊椎动物细胞和组织中发现的唾液酸鞘糖脂，发挥着广泛的作用。 在多种分子信号通路中的作用，这些信号通路影响人类疾病，包括糖尿病、癌症、神经- 退行性蛋白质病、智力残疾和许多其他疾病。神经节苷脂的主要结构为 定义明确的、有限的、在脊椎动物物种中共享的。它们的糖链不依赖于细胞信号调节 一种蛋白质载体。大多数神经节苷脂存在于细胞表面，其神经酰胺类脂包埋在 质膜及其多糖向外延伸。它们通过两种方式调节细胞生理，顺式和顺式。 互换。作为顺式调节因子，它们通过糖链与同一细胞中的跨膜蛋白侧向结合。 细胞膜来调节它们的功能。作为反式识别分子，它们与蛋白质在 细胞外环境或相反的细胞上，调节细胞与细胞之间的相互作用。顺式和反式交互都是 专用于神经节苷脂多糖结构，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与蛋白质的相互作用仍然知之甚少 可广泛使用的、适应性强的和经过验证的工具及其使用的优化方法。这就是我们的目标 解决这一需求的项目使用化学生物技术来合成一套定义的主要 神经节苷脂携带破坏性最小的双功能光反应和炔烃(点击化学)标签。 将生成优化和验证的协议，以将双功能标记的神经节苷脂传递到外部 神经节苷脂细胞质膜的小叶，大多数神经节苷脂在此居住和发挥作用。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，验证它们的使用 鉴定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 本项目期限内的生物医学研究实验室。可交付成果、神经节苷脂探针和 经过验证的使用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。