CAREER: Rare Sugar Activation and Incorporation into Bacterial Glycan Polymers

职业：稀有糖活化并掺入细菌聚糖聚合物

• 批准号: 2143170
• 负责人: Tania Lupoli
• 金额: $ 75万
• 依托单位: New York University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2143170&HistoricalAwards=false
• 关键词: CAREER; Rare; Sugar; Activation; Incorporation

With the support of the Chemistry of Life Processes (CLP) program in the Division of Chemistry, Tania Lupoli from New York University will investigate the synthesis and recognition of sugars unique to bacterial cell surfaces. Almost all bacteria contain polymers composed of sugars, called polysaccharides, that extend from their cell surfaces. Many bacterial surface polysaccharides contain rare sugars, which are bacteria-specific monosaccharides that are absent in mammals, including humans. While scientists recognize that rare sugars are important for mediating interactions between bacteria and their environments, there is still incomplete understanding of how rare sugar-containing polymers are assembled and how these enable survival in hosts during infection. The proposed experiments aim to develop chemical approaches to study enzymes that recognize rare sugars, with the goal of providing chemical tools to analyze bacterial interactions with their environments. This work will enable graduate students to obtain training in synthetic chemistry, biochemistry and microbiology. This project will also be integrated into educational outreach programs on infectious disease, bacterial survival and antibiotic design to inspire future scientists.This research project seeks to develop routes to nucleotide rare sugars, and to quantitatively characterize their interactions with glycosyltransferase enzymes that assemble bacterial surface polysaccharides. Rare bacterial sugars are over-represented in 6-deoxysugars, which lack the canonical hydroxyl group found on C6 of the hexose ring, and are typically L- as opposed to D-isomers. This proposal seeks to develop reagents to decipher the molecular consequences of installing sugars with uncommon stereochemistry that lack a 6-hydroxyl group into the cell surface of bacteria, with a focus on Gram-negative bacteria. Robust synthetic and chemoenzymatic routes to activated rare sugar substrates will be developed. The mechanism and catalytic sites of rare sugar-utilizing glycosyltransferases will be determined using detailed kinetic and mutagenesis studies. This work is expected to help establish molecular rules for recognition of 6-deoxysugars by bacterial glycosyltransferases that are currently uncharacterized.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系生命过程化学（CLP）项目的支持下，来自纽约大学的Tania Lupoli将研究细菌细胞表面特有糖的合成和识别。几乎所有的细菌都含有由糖组成的聚合物，称为多糖，从细胞表面延伸出来。许多细菌表面多糖含有稀有糖，这些糖是哺乳动物（包括人类）中不存在的细菌特异性单糖。 虽然科学家们认识到稀有糖对于介导细菌与其环境之间的相互作用非常重要，但对于稀有含糖聚合物如何组装以及这些聚合物如何在感染期间在宿主中存活仍然没有完全的了解。 拟议的实验旨在开发化学方法来研究识别稀有糖的酶，目的是提供化学工具来分析细菌与环境的相互作用。 这项工作将使研究生获得合成化学、生物化学和微生物学方面的培训。该项目还将整合到传染病、细菌存活和抗生素设计的教育推广计划中，以激励未来的科学家。该研究项目旨在开发核苷酸稀有糖的路线，并定量表征它们与组装细菌表面多糖的糖基转移酶的相互作用。罕见的细菌糖在6-脱氧糖中过度存在，其缺乏在己糖环的C6上发现的典型羟基，并且通常是L-而不是D-异构体。 该提案旨在开发试剂，以破译将具有不常见立体化学的糖（缺乏6-羟基）安装到细菌细胞表面的分子后果，重点是革兰氏阴性细菌。 将开发用于活化稀有糖底物的稳健的合成和化学酶促途径。 稀有糖利用糖基转移酶的机制和催化位点将使用详细的动力学和诱变研究来确定。 这项工作预计将有助于建立分子规则识别的6-脱氧糖的细菌糖基转移酶，目前uncharacterized。这一奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。