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

这一行动资助了NSF 2023财年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。该研究金支持一项针对该研究员的研究和培训计划，该计划将增加生物学中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创造更大、更复杂的结构，称为蛋白质，其中一些需要额外的改变，以确保正常的功能。在某些蛋白质中，一个这样的变化涉及到复杂的糖分子与特定氨基酸的结合；这个过程被称为糖基化。一旦连接在一起，这些糖就会发挥作用，控制蛋白质的功能以及蛋白质与其他分子的相互作用。如果没有糖基化，蛋白质就会不正常地发挥作用，扰乱基本的生物功能。在人类中，这可能会导致许多先天性疾病和癌症。尽管它发挥了关键作用，但人们对糖基化中使用的复杂糖是如何产生的知之甚少。因此，这项研究试图调查制造这些糖的酶；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，了解这些酶将创造新的见解，指导未来在生物技术中的应用，包括设计针对先天性糖基化障碍和癌症的新疗法。此外，这项工作将促进研究员直接培训和指导不同的受训者在他们自己的科学生涯中导航的能力。这项工作将通过定义磷酸转移酶和糖基转移酶的广泛催化机制，并制定适用于生命所有领域的基本原则，来丰富天冬酰胺连接的糖基化(NLG)的基础知识。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，这项工作将全面表征人磷酸转移酶DPATG1及其合作伙伴糖基转移酶ALG13和ALG14。这项工作还将研究其他重要的NLG糖基转移酶之间的新的相互作用。为了实现这一点，该研究员将通过冷冻EM确定人DPAGT1与新型抑制剂络合物的结构。然后，该研究员将对DPAGT1/ALG13/ALG14络合物进行结构表征。最后，该研究员将利用AlphaFold来探索未知的异寡聚复合体，并定义NLG途径中完整的膜磷酸转移酶和糖基转移酶的普遍机制。该研究员将参加低温电磁培训讲习班和特定领域的会议。最后，该研究员将在现有基础设施的基础上，为来自历史上被边缘化背景的学生和博士后创建一个校园范围的网络，寻求建立社区、专业发展，并将受训人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优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.