Artificial Glycosidase with Controlled Selectivity

具有受控选择性的人工糖苷酶

• 批准号: 10024717
• 负责人: Yan Zhao
• 金额: $ 29.91万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10024717
• 关键词: 3-Dimensional; Acidity; Active Sites; Affinity; Binding; Binding Sites; Biological; Biological Assay; Biological Process; Biology; Biomimetics; Buffers; Carbohydrates; Catalysis; Chemicals; Chemistry; Complement; Complex; Development; Endoglycosidases; Enzymes; Exoglycosidases; Generations; Genomics; Glycoside Hydrolases; Glycosides; Heterogeneity; Hydrolysis; Knowledge; Lectin; Length; Methods; Modification; Molecular; Oligosaccharides; Organic Synthesis; Oxygen; Peptides; Planet Earth; Polysaccharides; Positioning Attribute; Preparation; Proteins; Proteomics; Reporting; Research Design; Shapes; Site; Structure; Techniques; Technology; Time; Water; adduct; base; biological systems; catalyst; design; functional group; human disease; imprint; improved; insight; molecular recognition; molecular size; monomer; nanoparticle; public health relevance; receptor; reconstruction; small molecule; success; sugar; surfactant; three dimensional structure; tool

Artificial Glycosidase with Controlled Selectivity Abstract Carbohydrates are the most abundant biomolecules on the earth, and involved in numerous biological processes and all major human diseases. Glycoscience, nonetheless, lags behind genomics and proteomics, due to the extreme complexity, dynamic structural diversity, and micro-heterogeneity of glycans found in biological systems. Another reason, according to the 2012 NRC report “Transforming Glycoscience”, was the lack of suitable tools and methods “to detect, describe, and fully purify glycans…and then to characterize their chemical composition and structure. Molecular recognition of carbohydrates and peptides has been long-standing challenges in bioorganic and supramolecular chemistry, due to the importance of these molecules in biology. The PI’s group has developed protein-sized molecularly imprinted nanoparticles (MINPs) to bind a wide range of biologically interesting guests including carbohydrates and peptides. They are prepared and purified in < 2 days without any special techniques, once the template, functional monomers, and cross-linkable surfactants are available. MINP-based “synthetic lectins” were shown to recognize a wide range of mono- and oligosaccharides in water with tens of micromolar binding affinities. Oligosaccharides were distinguished based on their building blocks, glycosidic linkages, and chain length. The overall objective of this proposal is to develop synthetic glycosidases with selectivities unavailable in their natural counterparts. The proposed catalysts contain substrate-specific active sites with precisely installed catalytic groups for optimal catalysis. In the traditional synthesis of receptors and supramolecular catalysts, tremendous synthetic efforts are needed just to have a binding pocket. Fine tuning of the pocket for specific and complex biomolecules is nearly impossible. The micellar imprinting technology used in the MINP preparation, on the other hand, can quickly construct multifunctionalized, complex- shaped active sites from simple building blocks. The principles to be demonstrated are not limited to glycan hydrolysis and are expected to open up many possibilities in the design and synthesis of enzyme-mimicking catalysts.

一种选择性可控的人工糖苷酶 摘要 碳水化合物是地球上最丰富的生物分子，参与了 无数的生物过程和人类的所有重大疾病。然而，血糖科学， 落后于基因组学和蛋白质组学，由于结构极其复杂、动态 生物系统中发现的葡聚糖的多样性和微观异质性。另一个原因是， 根据NRC 2012年的报告《改造血糖科学》，是缺乏合适的 检测、描述和完全纯化多聚糖…的工具和方法然后要刻画 它们的化学成分和结构。 碳水化合物和多肽的分子识别由来已久。 生物有机化学和超分子化学的挑战，由于这些的重要性 生物学中的分子。Pi的团队已经开发出蛋白质大小的分子印迹 纳米颗粒(MINP)可结合多种具有生物意义的客体，包括 碳水化合物和多肽。它们是在&lt；2天内制备和纯化的，没有任何特殊的 技术，一旦模板、功能单体和可交叉连接的表面活性剂 可用。基于MINP的“合成凝集素”被证明能够识别广泛的单链凝集素。 低聚糖在水中具有数十个微摩尔结合亲和力。低聚糖 根据它们的组成单元、糖苷键和链长来区分它们。 这项提案的总体目标是开发具有以下特性的合成糖苷酶 它们的选择性是天然的同类所不具备的。建议的催化剂包括 具有精确安装的催化基团的底物特定活性部位，可实现最佳催化。 在传统的受体和超分子催化剂的合成中，大量的合成 仅仅是为了有一个有约束力的口袋就需要努力。对口袋进行微调，以实现特定和 复杂的生物分子几乎是不可能的。胶束印迹技术在电子显微镜中的应用 另一方面，MINP制剂可以快速构建多功能、复杂的 从简单的构建块塑造活跃的站点。要论证的原则不是 仅限于葡聚糖的水解，预计将在设计中开辟许多可能性 以及仿酶催化剂的合成。