NSF Postdoctoral Fellowship in Biology: Illuminating mechanisms of essential asparagine-linked glycosylation enzymes

NSF 生物学博士后奖学金：阐明必需天冬酰胺连接糖基化酶的机制

• 批准号: 2305964
• 负责人: Jessica Ochoa
• 金额: $ 24万
• 依托单位: Ochoa, Jessica Micaela
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2305964&HistoricalAwards=false
• 关键词: NSF; Postdoctoral; Fellowship; Biology; Illuminating

This action funds an NSF Postdoctoral Research Fellowship in Biology for FY 2023, Broadening Participation of Groups Underrepresented in Biology. The Fellowship supports a research and training plan for the Fellow that will increase the participation of groups underrepresented in biology. Amino acids are one of the essential building blocks of life. All cells use amino acids to create larger, more complex structures called proteins, with several requiring additional changes to ensure proper function. In some proteins, one such change involves the attachment of complex sugar molecules to specific amino acids; this process is called glycosylation. Once attached, these sugars play roles in controlling how proteins function and how proteins interact with other molecules. Without glycosylation, proteins would function improperly, disrupting essential biological functions. In humans, this can lead to numerous congenital disorders and cancers. Despite its critical role, little is understood about how the complex sugars used in glycosylation are generated. Thus, this research seeks to investigate the enzymes that make these sugars; this work explores enzyme function by understanding their molecular structures. Because glycosylation is important for all life, understanding these enzymes will create new insights that guide future applications in biotechnology, including designing new therapeutics that target congenital disorders of glycosylation and cancers. Moreover, this work will facilitate the fellow’s ability to directly train and mentor diverse trainees navigating their own scientific careers. This work will enrich foundational knowledge of asparagine-linked glycosylation (NLG) by defining broad catalytic mechanisms of phosphotransferases and glycosyltransferases, and formulating underlying principles applicable to NLG in all domains of life. Using recombinant protein expression in human cells, structural biology, bioinformatics, and biochemistry, this work will fully characterize the human phosphotransferase, DPATG1, and its partner glycosyltransferases Alg13 and Alg14. This work will also investigate novel interactions between other essential NLG glycosyltransferases. To achieve this, the fellow will determine the structure of human DPAGT1 in complex with novel inhibitors by cryo-EM. Then, the fellow will structurally characterize the DPAGT1/Alg13/Alg14 complex. Lastly, the fellow will leverage AlphaFold to probe uncharacterized heterooligomeric complexes and define universal mechanisms of integral membrane phosphotransferases and glycosyltransferases from the NLG pathway. The fellow will attend cryo-EM training workshops and field-specific conferences. Lastly, the fellow will build upon existing infrastructure to create a campus-wide network for students and postdocs from historically marginalized backgrounds that seeks to build community, professional development, and connect trainees with existing resources.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.

这项行动资助了2023财年的NSF生物学博士后研究奖学金，扩大了生物学中代表性不足的群体的参与。该研究金支持研究员的一项研究和培训计划，该计划将增加在生物学领域代表性不足的群体的参与。氨基酸是生命的基本组成部分之一。 所有细胞都使用氨基酸来产生更大，更复杂的结构，称为蛋白质，其中一些需要额外的变化以确保正常功能。 在某些蛋白质中，一种这样的变化涉及将复杂的糖分子连接到特定的氨基酸上;这个过程称为糖基化。 一旦附着，这些糖在控制蛋白质如何发挥功能以及蛋白质如何与其他分子相互作用方面发挥作用。 如果没有糖基化，蛋白质将不能正常发挥功能，破坏基本的生物功能。 在人类中，这可能导致许多先天性疾病和癌症。 尽管它的关键作用，很少有人了解如何在糖基化中使用的复杂糖产生。 因此，这项研究旨在研究制造这些糖的酶;这项工作通过了解它们的分子结构来探索酶的功能。 由于糖基化对所有生命都很重要，了解这些酶将产生新的见解，指导未来的生物技术应用，包括设计针对先天性糖基化疾病和癌症的新疗法。 此外，这项工作将促进研究员的能力，直接培训和指导不同的学员导航自己的科学生涯。这项工作将丰富基础知识的天冬酰胺连接的糖基化（NLG）通过定义广泛的催化机制的磷酸转移酶和糖基转移酶，并制定基本原则适用于NLG在所有领域的生活。 利用重组蛋白在人类细胞中的表达，结构生物学，生物信息学和生物化学，这项工作将充分表征人类磷酸转移酶，DPATG 1，及其伴侣糖基转移酶Alg 13和Alg 14。 这项工作还将研究其他必需的NLG糖基转移酶之间的新的相互作用。 为了实现这一目标，该研究员将通过cryo-EM确定与新型抑制剂复合的人DPAGT 1的结构。 然后，研究员将对DPAGT 1/Alg 13/Alg 14复合物进行结构表征。 最后，该研究员将利用AlphaFold来探测未表征的异源寡聚复合物，并定义来自NLG途径的完整膜磷酸转移酶和糖基转移酶的通用机制。 该研究员将参加低温电磁培训讲习班和具体领域的会议。 最后，该研究员将建立在现有的基础设施，为来自历史边缘背景的学生和博士后创建一个校园范围的网络，旨在建立社区，专业发展，并将学员与现有资源联系起来。该奖项反映了NSF的法定使命，并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，被认为值得支持。