Molecular mechanisms of membrane protein homeostasis at the Golgi

高尔基体膜蛋白稳态的分子机制

• 批准号: 10713404
• 负责人: Richa Sardana
• 金额: $ 37.57万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713404
• 关键词: Anabolism; Cells; Cellular biology; Congenital disorders of glycosylation; Defect; Disease; Enzymes; Eukaryotic Cell; Flow Cytometry; Functional disorder; Genetic; Glycoconjugates; Glycolipids; Glycoproteins; Goals; Golgi Apparatus; Health; Homeostasis; Human; Image; Immune System Diseases; In Vitro; Knowledge; Life; Location; Malignant Neoplasms; Maps; Membrane; Membrane Proteins; Membrane Transport Proteins; Molecular; Pathway interactions; Play; Process; Proteins; Proteomics; Recycling; Role; Toxin; Virus; experimental study; glycosyltransferase; human disease; interest; new therapeutic target; novel; programs; proteostasis; receptor; receptor recycling; reconstitution; solute; sugar nucleotide; trafficking; vesicle transport

Project summary/Abstract The Golgi complex plays a prominent role in secretory and endocytic trafficking in eukaryotic cells and is key to the biosynthesis of glycoconjugates (glycoproteins and glycolipids) that are essential for life. Golgi resident proteins, such as glycosyltransferases and sugar nucleotide transporters, are precisely distributed across the Golgi stacks by recycling mechanisms that counteract the flow of ongoing vesicular transport. Dysfunction in these mechanisms, or their hijacking by viruses and toxins, is known to have serious consequences for human health, leading to congenital disorders of glycosylation, cancers, and immune dysfunction. Membrane proteins residing in various Golgi compartments are well annotated; however, the mecha- nistic basis of how most Golgi proteins are selected for recycling, or how these processes are regulated are poorly understood. My lab is interested in uncovering these fundamental mechanisms governing Golgi homeostasis using a multifaceted approach combining genetics, flow-cytometry, imaging, in vitro reconstitution, and proteomics. In preliminary results, we have identified novel transmembrane compo- nents orchestrating recycling of specific subsets of Golgi enzymes. Our findings have opened doors for interrogating new players and dissecting the mechanisms critical to maintain Golgi identity and function. Over the next five years, our goals are to (1) identify novel recycling receptors required at different Golgi compartments and establish a systematic map of the intra-Golgi recycling network, (2) determine how the transmembrane receptors engage with their cargos, and (3) define the novel functions of a disease- associated membrane transporter in solute transport and protein recycling in the Golgi. The combined results of our experiments will elucidate how multiple recycling pathways sustain normal Golgi function, and how this homeostasis is disrupted in human disease.

项目概要/摘要 高尔基复合体在真核细胞的分泌和内吞运输中起着重要作用 并且是糖缀合物（糖蛋白和糖脂）的生物合成的关键，糖缀合物对于 生活高尔基体的常驻蛋白，如糖基转移酶和糖核苷酸转运蛋白， 通过循环机制分布在高尔基体上，这种机制抵消了正在进行的囊泡的流动， 运输这些机制的功能障碍，或它们被病毒和毒素劫持， 对人类健康造成严重后果，导致先天性糖基化障碍、癌症和 免疫功能紊乱 存在于各种高尔基体隔室中的膜蛋白被很好地注释;然而， 大多数高尔基体蛋白如何被选择用于再循环，或这些过程如何被调节的物理基础 我们对此知之甚少。我的实验室有兴趣揭示这些基本的机制， 结合遗传学、流式细胞术、成像、体外等多方面方法的高尔基体内平衡 重组和蛋白质组学。在初步结果中，我们已经确定了新的跨膜化合物， 协调高尔基体酶的特定子集的再循环的元素。我们的发现为 询问新的参与者，剖析维持高尔基体身份和功能的关键机制。 在接下来的五年里，我们的目标是：（1）确定不同高尔基体所需的新型回收受体 隔室，并建立一个系统的地图内高尔基体回收网络，（2）确定如何 跨膜受体与它们的货物结合，和（3）定义疾病的新功能- 在高尔基体的溶质运输和蛋白质再循环中与膜转运蛋白相关。将合并的 我们的实验结果将阐明多种循环途径如何维持正常的高尔基体功能， 以及这种体内平衡在人类疾病中是如何被破坏的。