Deciphering molecular details of cellular sugar transport and their roles in disease

破译细胞糖转运的分子细节及其在疾病中的作用

• 批准号: 10799018
• 负责人: Jeffrey S Abramson
• 金额: $ 3.9万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10799018
• 关键词: Cells; Citric Acid Cycle; Collaborations; Complement; Cryoelectron Microscopy; Data; Disease; Family; Glucose; Golgi Apparatus; Human; Ion Channel; Lipid Bilayers; Lipids; Mediator; Membrane Transport Proteins; Metabolic; Mitochondria; Molecular; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Organelles; Organism; Outer Mitochondrial Membrane; Parents; Pathway interactions; Performance; Pharmacologic Substance; Physiology; Process; Production; Protein Glycosylation; Protein Isoforms; Proteins; Resolution; Resources; Role; Sialic Acids; Site; Sodium; Source; Structure; Therapeutic; Voltage-Dependent Anion Channel; X ray diffraction analysis; inhibitor; pathogen; sialic acid permease; small molecule; structural biology; sugar; sugar nucleotide; symporter; virtual

Abstract Sugars, in particular glucose, are not only a ubiquitous cellular fuel source in virtually all organisms, but also serve as critical metabolic intermediates in which activated glucose molecules are transported to the ER and Golgi and used for glycosylating proteins, lipids and other organic compounds as part of the biosynthetic-secretory pathway. To accomplish these diverse and localized functions, the body utilizes membrane transporters and channels to transfer glucose and its intermediates across the otherwise impermeable membrane lipid bilayer that surrounds all cells and organelles. Secondary active transporters are key mediators in this process. Alternatively, the Voltage Dependent Anion Channel funnels glucose intermediates into the mitochondria where they enter the TCA cycle for the production of ATP. In their essential function for physiology, these proteins are implicated in numerous diseases and are designated targets for pharmaceutical compounds. In the parent R35 entitled, “Deciphering molecular details of cellular sugar transport and their roles in disease” we aim to characterize the structure and function of several families of transporters involved in cellular sugar and metabolite transport. Specifically, we want to study the Sodium Glucose Cotransporter (SGLT), Nucleotide Sugar Transporters (NST) and Sialic Acid Transporter (SiaT) and the Voltage-Dependent Anion Channel (VDAC). Human SGLTs are well known targets for treating Type II diabetes, but the molecular details of inhibition and the functional differences between isoforms are not well understood. This is in large part due to the lack of structural information. NSTs import various activated sugar compounds into the Golgi and ER, whereas SiaT serves to import scavenged sialic acids from its host. Differences between eukaryotic and pathogen NSTs could be exploited for therapeutic purposes. However, this family of transporters is still largely uncharacterized. VDAC is the central mediator of metabolite exchange through the outer mitochondrial membrane. Despite this critical role, key aspects of its functional gating and substrate transport are not well understood. Again, we hope that resolving additional structures of VDAC will help to answer these remaining questions. For all structure determination projects we need to collect, store and process large amounts of data. Our structural biology approach is complemented by functional studies to obtain a complete picture of sugar transport at an atomic resolution. We need to assess the functionality of heterologously- expressed proteins during the early stages of project. After obtaining the structures, we plan to interogate them by inserting mutations at critical sites identified in the structures and screen small- molecules as potential inhibitors.

抽象的 糖，特别是葡萄糖，不仅是几乎所有生物体中普遍存在的细胞燃料来源，而且 还充当关键的代谢中间体，其中活化的葡萄糖分子被转运至 内质网和高尔基体，用于糖基化蛋白质、脂质和其他有机化合物，作为 为了完成这些多样化和局部的功能，身体。 利用膜转运蛋白和通道将葡萄糖及其中间体转运穿过 围绕所有细胞和细胞器的其他不可渗透的膜脂质双层。 活性转运蛋白是该过程中的关键介体，或者电压依赖性阴离子。 通道将葡萄糖中间体输送到线粒体中，在那里它们进入 TCA 循环 在其生理学的基本功能中，这些蛋白质与许多相关因素有关。 疾病并被指定为药物化合物的靶点。 在母 R35 中，标题为“破译细胞糖转运的分子细节及其 在疾病中的作用”我们的目标是描述几个转运蛋白家族的结构和功能 具体来说，我们想研究葡萄糖钠。 协同转运蛋白 (SGLT)、核苷酸糖转运蛋白 (NST) 和唾液酸转运蛋白 (SiaT) 以及 电压依赖性阴离子通道 (VDAC) 是众所周知的治疗类型的靶点。 II 型糖尿病，但抑制的分子细节和异构体之间的功能差异是 这在很大程度上是由于缺乏 NST 导入的各种结构信息。 活化的糖化合物进入高尔基体和内质网，而 SiaT 则负责输入清除的唾液酸 真核生物和病原体 NST 之间的差异可用于研究。 然而，该转运蛋白家族在很大程度上仍未被表征。 尽管如此，它还是通过线粒体外膜进行代谢物交换的中心介质。 同样，其功能门控和底物传输的关键作用、关键方面尚不清楚。 我们希望解决 VDAC 的其他结构将有助于回答这些剩余的问题。 对于所有结构确定项目，我们都需要收集、存储和处理大量数据。 我们的结构生物学方法辅以功能研究，以获得完整的图景 我们需要评估异源的功能。 在项目的早期阶段表达的蛋白质在获得结构后，我们计划 通过在结构中确定的关键位点插入突变来询问它们，并筛选小 分子作为潜在的抑制剂。