N-glycosylation mechanism in insect cells.

昆虫细胞中的 N-糖基化机制。

• 批准号: 6929722
• 负责人: Donald L. Jarvis
• 金额: $ 23.35万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6929722
• 关键词: Baculoviridae; Insecta; SDS polyacrylamide gel electrophoresis; Sf9 cell line; animal tissue; biotechnology; enzyme activity; galactosyltransferases; gene expression; glycoprotein biosynthesis; glycoprotein structure; glycosylation; immunoelectron microscopy; immunoprecipitation; insect virus; mannosidase; oligosaccharides; polymerase chain reaction; protein engineering; protein structure function; recombinant proteins; tissue /cell culture; transfection /expression vector; transferase; virus diseases; western blottings

DESCRIPTION (provided by the applicant): Insect cells are widely known as hosts for baculovirus expression vectors (BEVs), which can produce large amounts of recombinant proteins with eukaryotic modifications. The insect cell-BEV system has been used to produce many different recombinant proteins, facilitating biomedical research on their structures, functions, and roles in disease. Many companies are using the insect cell-BEV system to produce recombinant proteins for high-throughput functional screens and to develop vaccines, therapeutics, and diagnostic reagents. The advantages of using insect cells for these applications include ease of scale-up, lower costs, and the absence of human adventitious agents. These important research activities require a dear understanding of protein biosynthesis and processing pathways in insect cells. However, this has not been a major topic of research. This proposal focuses on the insect cell protein N-glycosylation pathway. The long-term goals are to provide an unequivocal and detailed view of this important metabolic pathway in insect cells and to use this knowledge to develop new insect-based protein production systems with more extensive recombinant glycoprotein processing capabilities. The widespread use of insect cells as protein production systems for many important biomedical research projects is a strong justification for this research. Intelligent use of insect cells for recombinant glycoprotein production will require a better understanding of their N-glycosylation pathway. This understanding can direct innovative metabolic engineering efforts to produce new insect ceft-BEV systems with improved N-glycoprotein processing capabilities. Insect cells also occupy an important niche as models for fundamental research in the emerging field of glycobiology. The insect cell N-glycosylation pathway lies in-between the pathways of lower and higher eukaryotes in complexity. Thus, well-defined differences in insect protein glycosylation pathways could be exploited as specific targets to develop future insecticides. This could lead to better ways to control agriculturally and medically important insects and the diseases they help to spread, which would have a major impact on public health, worldwide.

描述（由申请人提供）：昆虫细胞被广泛认为是杆状病毒表达载体（BEV）的宿主，其可以产生大量具有真核修饰的重组蛋白。昆虫细胞-BEV系统已被用来生产许多不同的重组蛋白，促进对其结构、功能和在疾病中的作用的生物医学研究。许多公司正在使用昆虫细胞-BEV 系统生产用于高通量功能筛选的重组蛋白，并开发疫苗、治疗剂和诊断试剂。在这些应用中使用昆虫细胞的优点包括易于扩大规模、成本较低以及不存在人类外来因子。这些重要的研究活动需要对昆虫细胞中的蛋白质生物合成和加工途径有深入的了解。然而，这并不是研究的主要课题。该提案重点关注昆虫细胞蛋白 N-糖基化途径。长期目标是提供昆虫细胞中这一重要代谢途径的明确而详细的视图，并利用这些知识开发具有更广泛的重组糖蛋白加工能力的新的基于昆虫的蛋白质生产系统。 昆虫细胞作为许多重要生物医学研究项目的蛋白质生产系统的广泛使用是这项研究的有力理由。智能地利用昆虫细胞进行重组糖蛋白生产需要更好地了解其 N-糖基化途径。这种理解可以指导创新的代谢工程工作，以生产具有改进的 N-糖蛋白加工能力的新型昆虫 ceft-BEV 系统。昆虫细胞作为新兴糖生物学领域基础研究的模型也占据着重要的地位。昆虫细胞 N-糖基化途径的复杂性介于低等真核生物和高等真核生物的途径之间。因此，昆虫蛋白糖基化途径的明确差异可以作为开发未来杀虫剂的具体目标。这可能会带来更好的方法来控制农业和医学上重要的昆虫及其传播的疾病，这将对全世界的公共卫生产生重大影响。