Facile Synthesis of O-Glycans and O-Glycopeptides

O-聚糖和 O-糖肽的简便合成

• 批准号: 8985647
• 负责人: Lei Li
• 金额: $ 49.3万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-05 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8985647
• 关键词: Amino Acids; Anabolism; Automation; Chemicals; Complex; DNA; Data; Databases; Development; Dreams; Enzymes; Glycoconjugates; Glycopeptides; Human; Libraries; Marketing; Methodology; N-acetylglucopyranosylamine; Oligosaccharides; Pathway interactions; Peptides; Phase; Polymers; Polysaccharides; Production; Protocols documentation; Publications; Reaction; Research; Research Contracts; Route; S Phase; Scientist; Solid; Structure; System; Technology; Text; Training; Update; Validation; Work; abstracting; base; catalyst; chemical synthesis; cost; expression cloning; glycosylation; glycosyltransferase; man; meetings; milligram; podoplanin; programs; research study; success; sugar; web site

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. This proposal is developing core synthesis – enzymatic extension (CSEE) approach to produce large glycans and glycopeptides with most natural structural diversity. In CSEE approach, a core comprising a few sugars in the reducing end is synthesized first. Convergent core synthesis is cost-efficient with well documented methodologies. Then glycosyltransferases are used to elongate the core by following a variety of different biosynthesis pathways to generate complex and larger glycoconjugates with high diversity. Depending on high region- and stereo-selectivity of glycosyltransferases, CSEE is the most efficient approach to produce complex glycoconjugates with high fidelity. Our recent success of using 7 chemically synthesized N-glycan cores and 4 glycosyltransferases to produce 73 complex N-glycans clearly demonstrates that CSEE is an answer to the complexity and diversity of glycomes. In this program, 8 cores of O-GalNAc-Ser/Thr will be synthesized and extended with 16 glycosyltransferases to afford focused and diverse O-GalNAc-glycans/glycopeptides libraries; 3 cores of O-Man-Ser/Thr will be synthesized and extended with 9 glycosyltransferase to produce O-Man glycans/glycopeptides; O-Fuc, O-Glc and O-Xyl glycans and glycopeptides will be produced similarly. CSEE for synthesis of these O-glycan/O-glycopeptides will be fully validated in other labs. The technology advance of this proposal is to automate the enzymatic extension by using a conventional peptide synthesizer. We will fully investigate stability and optimal reaction conditions for the most synthetic useful glycosyltransferases and make the information accessible via a website. In summary, this program should make such a dream come true: scientists can have affordable access to any N- & O-glycans and glycopeptides as they have for regular peptides.

在此输入文本，它是应用程序的新摘要信息。本节必须是否 超过30行的文本。 该提案正在开发核心合成-酶促延伸（CSEE）方法， 具有最天然结构多样性的大聚糖和糖肽。在CSEE方法中， 在还原端包含少量糖的糖。收敛核合成是 具有成本效益，方法有据可查。然后糖基转移酶用于 通过遵循各种不同的生物合成途径来延长核心以产生复合物 和具有高多样性的较大的糖缀合物。取决于高区域和立体选择性 在糖基转移酶中，CSEE是生产复杂糖缀合物的最有效方法 高保真度。我们最近使用7个化学合成的N-聚糖核心和4个 糖基转移酶产生73个复杂的N-聚糖清楚地表明，CSEE是一种 解决了糖组的复杂性和多样性。 在本计划中，将合成8个O-GalNAc-Ser/Thr核心，并将其延伸为16个 糖基转移酶，以提供集中和多样化的O-GalNAc-聚糖/糖肽文库; 3 O-Man-Ser/Thr的核心将被合成并用9糖基转移酶延伸以产生 O-Man聚糖/糖肽; O-Fuc、O-Glc和O-Xyl聚糖和糖肽将 制作类似。将对合成这些O-聚糖/O-糖肽的CSEE进行充分验证 在其他实验室。该建议的技术进步是通过以下方式使酶促延伸自动化： 使用常规的肽合成仪。我们将充分研究稳定性和最佳反应 最有用的合成糖基转移酶的条件，并使信息可访问 通过网站。 总之，这个计划应该让这样一个梦想成真：科学家可以有负担得起的 获得任何N-和O-聚糖和糖肽，因为它们具有常规肽。