Contribution of N-glycosylation to the Toxoplasma glycoproteome

N-糖基化对弓形虫糖蛋白质组的贡献

• 批准号: 8432797
• 负责人: ANTHONY P. SINAI
• 金额: $ 18.02万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8432797
• 关键词: Acquired Immunodeficiency Syndrome; Acute; Address; Affinity; Affinity Chromatography; Animals; Apicomplexa; Area; Asparagine; Bioinformatics; Biological Process; Biology; Blood; Brain; Cataloging; Catalogs; Cells; Chronic; Chronic Phase; Complex; Computer Simulation; Coupled; Cyst; Development; Disease; Elements; Enzymes; Exhibits; Foundations; Funding Mechanisms; Future; Gene Expression; Genes; Genetic; Glycoproteins; Golgi Apparatus; HIV; Homologous Gene; Immune; Immune response; Immune system; Incidence; Individual; Infection; Investigation; Knock-out; Knowledge; Lectin; Life; Light; Link; Maintenance; Mass Spectrum Analysis; Mediating; Methods; Muscle; Nature; Neospora; Opportunistic Infections; Organelles; Parasites; Pathogenesis; Pathology; Pathway interactions; Patients; Pattern; Phase; Play; Polysaccharides; Protein Glycosylation; Proteins; Proteome; Proteomics; Protozoa; Role; Seroprevalences; Site-Directed Mutagenesis; Sterility; Structure; System; Technology; Testing; Tetanus Helper Peptide; Tissues; Toxoplasma; Toxoplasma gondii; Vision; Work; asexual; base; genetic manipulation; genome analysis; glycosylation; high risk; infancy; knowledge base; member; microbial; rhoptry; sugar

DESCRIPTION (provided by applicant): The protozoan parasite Toxoplasma gondii is an important opportunistic infection causing life threatening disease in patients with HIV-AIDS and other immune compromising conditions. The high seroprevalence of the T. gondii worldwide but low incidence of symptomatic disease reflects the fact that the acute phase of infection is effectively handled by the immune response. Unfortunately a sterile cure is not achieved. Rather the parasite establishes a life-long chronic infections mediated by tissue cysts typically formed in the brain and muscle. The tissue cyst is protected by a heavily glycosylated cyst wall. Despite the importance of glycosylation, little is known about these pathways in the parasite. With this work we address the contribution of N-linked glycosylation in Toxoplasma. We propose to use lectin affinity coupled with mass spectrometric identification to catalogue the diversity of N-glycosylated glycoproteins in tissue cysts. We further plan to conditionally knock out a key gene in the N-glycosylation, TgALG7, in order to assess its role in parasite biology with a focus on protein targeting to its diverse unique secretory organelles. Given the central role for glycosylation in the tissue cyst we propose an experimental strategy to selectively knock out gene expression in the tissue cyst. This technology will be applied to establishing the contribution of TgALG7 and by extension N-glycosylation to tissue cyst formation and stability. This work represents the first in depth study of the cell biologic basis of protein glycosylation i Toxoplasma using state of the art approaches to modulate gene expression.

描述（由申请人提供）：原生动物寄生虫弓形虫弓形虫是一种重要的机会性感染，导致HIV-AID和其他免疫损害疾病患者的生命危险疾病。全世界的T. gondii的高血清阳性率高但有症状疾病的发病率低，反映了一个事实，即免疫反应有效地处理了感染的急性阶段。不幸的是，没有实现无菌治疗。相反，寄生虫建立了终身慢性感染，该感染是由通常在大脑和肌肉中形成的组织囊肿介导的。组织囊肿受到严重的糖基化囊肿壁的保护。尽管糖基化的重要性很重要，但对寄生虫中的这些途径知之甚少。通过这项工作，我们解决了N-连接的糖基化在弓形虫中的贡献。我们建议使用凝集素亲和力，再加上质谱鉴定来分类组织囊肿中N-糖基化糖蛋白的多样性。我们进一步计划将N-糖基化TGALG7中的一个关键基因有条件地淘汰，以评估其在寄生虫生物学中的作用，重点是靶向其多样化的独特分泌细胞器。鉴于糖基在组织囊肿中的核心作用，我们提出了一种实验策略，可以选择性地敲除组织囊肿中的基因表达。该技术将应用于建立TGALG7的贡献，并通过扩展N-糖基化对组织囊肿形成和稳定性的贡献。这项工作代表了使用最终方法调节基因表达的最深入研究蛋白质糖基化I毒品的细胞生物学基础。