Structure and Function of the Circumsporozoite Protein

环子孢子蛋白的结构和功能

• 批准号: 8037654
• 负责人: Photini Sinnis
• 金额: $ 20.7万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8037654
• 关键词: Adhesions; Adhesives; Adult; Antigens; Antimalarials; Architecture; Behavior; Binding; Biochemical; Biological Assay; Blood; Bypass; C-terminal; Cell surface; Cells; Child; Cleaved cell; Communicable Diseases; Complex; Confocal Microscopy; Culicidae; Data; Databases; Dermis; Development; Drug resistance; Effector Cell; Event; Excision; Future; Gene Expression Profile; Generations; Genes; Goals; Heparin Binding; Hepatocyte; Immune; Immune Sera; Infection; Life; Link; Liver; Location; Maintenance; Malaria; Malaria Vaccines; Malaria prevention; Masks; Membrane Proteins; Methods; Midgut; Modeling; Molecular Conformation; Mutagenesis; N-terminal; Organ; Parasites; Peptide Hydrolases; Pharmacotherapy; Phase; Phenotype; Plasmodium; Prevention; Process; Property; Protease Gene; Proteins; Proteolysis; Proteolytic Processing; Proteome; Recombinant Proteins; Role; Route; Salivary Glands; Site; Skin; Sporozoites; Staging; Structure; Surface; System; Techniques; Tertiary Protein Structure; Travel; Vaccines; Work; base; circumsporozoite protein; design; improved; migration; mutant; protein function; public health relevance; research study; success; transmission process; vaccine candidate; vaccine development

DESCRIPTION (provided by applicant): Plasmodium sporozoites make a remarkable journey from the mosquito midgut wall to the mammalian liver. It is the overall goal of this proposal to elucidate the role of the sporozoite's major surface protein, the circumsporozoite protein (CSP) in this journey. We have been studying the proteolytic processing of CSP and thus far, have found that processing occurs when sporozoites contact hepatocytes and is required for efficient invasion. Specifically, our data indicate that the amino-terminus of CSP masks the cell-adhesive domain in the carboxy-terminus and that proteolytic cleavage results in removal of the amino-terminus and exposure of this cell-adhesive domain. Overall our data suggest that CSP has two functional domains, each with distinct role(s) during the sporozoite's journey. The goal of this proposal is to continue our structure-function work on this important protein to define the role of each CSP domain in the mammalian host and to identify the protease responsible for CSP cleavage. The specific aims are: 1) To elucidate the phenotype of sporozoite mutants which expressed only the cleaved form of CSP in the dermis of the mammalian host. Using quantitative PCR, transcriptional profiling and confocal microscopy, we will determine whether N-terminal deletion mutants remain in the skin and begin their development there. 2) To determine the binding properties of each CSP domain by generating mutants in N-terminal residues that we have identified as having potential heparin-binding activity and using biochemical techniques to determine the binding partner of the TSR domain. 3) To identify the protease that cleaves CSP and determines its subcellular location in the sporozoite. These studies will be performed using conditional mutagenesis to delete candidate protease genes and using antisera to localize the proteases in sporozoites. Overall these studies will elucidate how CSP functions in the sporozoite's journey from mosquito to mammalian host and should open up new avenues for the control and prevention of malaria. PUBLIC HEALTH RELEVANCE: These studies are of relevance to the control and prevention of one of the most important infectious diseases in the world. Malaria continues to impact significantly, children and adults living in tropical and subtropical parts of the world, in part due to drug resistance to our best anti-malarials and to the lack of an effective vaccine. The experiments described in this proposal could provide the basis for new drug therapies and improve upon existing approaches to malaria vaccine development.

描述（由申请人提供）：疟原虫子孢子从蚊子中肠壁到哺乳动物肝脏进行了一次非凡的旅程。阐明子孢子的主要表面蛋白环子孢子蛋白（CSP）在这一过程中的作用是本提案的总体目标。我们一直在研究CSP的蛋白水解加工，到目前为止，已经发现加工发生在子孢子接触肝细胞时，并且是有效侵入所必需的。具体而言，我们的数据表明，CSP的氨基末端掩盖了羧基末端的细胞粘附结构域，蛋白水解切割导致氨基末端的去除和该细胞粘附结构域的暴露。总的来说，我们的数据表明，CSP有两个功能域，每一个都有不同的作用（S）在子孢子的旅程。该提案的目标是继续我们对这一重要蛋白质的结构-功能研究，以确定每个CSP结构域在哺乳动物宿主中的作用，并确定负责CSP切割的蛋白酶。具体目标是：1）阐明在哺乳动物宿主真皮中仅表达CSP裂解形式的子孢子突变体的表型。使用定量PCR，转录谱和共聚焦显微镜，我们将确定是否N-末端缺失突变体留在皮肤和开始他们的发展有。2)通过在我们已确定具有潜在肝素结合活性的N-末端残基中产生突变体，并使用生物化学技术确定TSR结构域的结合伴侣，确定每个CSP结构域的结合特性。3)目的鉴定切割CSP的蛋白酶，并确定CSP在子孢子中的亚细胞定位。这些研究将使用条件诱变来删除候选蛋白酶基因，并使用抗血清来定位子孢子中的蛋白酶。总的来说，这些研究将阐明CSP如何在子孢子从蚊子到哺乳动物宿主的旅程中发挥作用，并为疟疾的控制和预防开辟新的途径。 公共卫生相关性：这些研究与世界上最重要的传染病之一的控制和预防相关。疟疾继续严重影响生活在世界热带和亚热带地区的儿童和成人，部分原因是对我们最好的抗疟疾药物的耐药性以及缺乏有效的疫苗。该提案中描述的实验可以为新的药物疗法提供基础，并改进现有的疟疾疫苗开发方法。