Molecular characterization and substrate identification of malaria kinase PfCDPK5

疟疾激酶 PfCDPK5 的分子表征和底物鉴定

• 批准号: 8525534
• 负责人: JEFFREY D DVORIN
• 金额: $ 43.79万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-15 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8525534
• 关键词: Age-Years; Antimalarials; Binding; Biological Process; Blood; Calcium; Calcium Binding; Candidate Disease Gene; Cells; Cessation of life; Child; Child Mortality; Clinical; Developing Countries; Development; Disease; Epitopes; Erythrocytes; Essential Genes; Evaluation; Future; Genetic; Goals; Human; Human Genome; In Vitro; Infection; Investigation; Life; Life Cycle Stages; Longitudinal Studies; Malaria; Mediator of activation protein; Molecular; Molecular Genetics; Morbidity - disease rate; Parasites; Peptide Hydrolases; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Plasmodium; Plasmodium falciparum; Pregnant Women; Process; Proteins; Proteomics; Public Health; Recombinants; Regulation; Resistance; Resources; Role; Signal Pathway; Signal Transduction; Staging; System; Tars; Techniques; Tertiary Protein Structure; Therapeutic; Time; Toxoplasma gondii; Transgenic Organisms; Vaccines; Yeasts; analog; asexual; base; burden of illness; calcium-dependent protein kinase; combat; design; falls; genetic analysis; knock-down; member; mortality; mutant; novel; prevent; public health relevance; tool; vector control; yeast two hybrid system

DESCRIPTION (provided by applicant): Malaria is among the leading causes of mortality for children under five years of age worldwide, with most of these deaths resulting from Plasmodium falciparum infection. Resistance to existing anti-malarial medications is an urgent problem and may prevent effective eradication strategies. A molecular understanding of the life cycle of P. falciparum will facilitate the rational design of new therapies. Efficient egress of P. falciparum out of an infected human red blood cell is a fundamental step in the parasite life cycle, and a step that is not targeted by current anti-malarial therapeutics. PfCDPK5 has recently been identified as a kinase that is critical for parasite egress. Using a state-of-the-art inducible protein destabilization system, functional evaluation of this essential gene is now possible. Transgenic parasites with an inducible knockdown in PfCDPK5 are arrested prior to egress. The ability to regulate the level of PfCDPK5 provides a unique resource to study both the mechanism of activation and the downstream effectors of this essential kinase. We hypothesize that PfCDPK5 is a central mediator of the calcium-based egress signaling pathway. The immediate goals of this proposal are to gain a molecular understanding of PfCDPK5 activation and characterize its role in the egress signaling pathway. In the first aim, a molecular genetic analysis of PfCDPK5 activation will be conducted using transgenic parasites that allow inducible regulation of essential protein levels. In the second aim, the downstream substrate(s) of PfCDPK5 will be discovered using both a candidate gene approach as well as advanced proteomic techniques. The long- term objectives and public health implications of these studies are to identify novel targets for new anti- malarial therapeutics. This long-term goal will be achieved as a direct result from the molecular characterization of the essential PfCDPK5 signaling pathway in P. falciparum parasites.

描述（由申请人提供）：疟疾是全世界五岁以下儿童死亡的主要原因之一，其中大多数死亡是由恶性疟原虫感染引起的。对现有抗疟疾药物的耐药性是一个紧迫的问题，可能会阻碍有效的根除策略。对恶性疟原虫生命周期的分子理解将有助于新疗法的合理设计。 P 的有效出口。 从受感染的人类红细胞中分离出恶性疟原虫是寄生虫生命周期的一个基本步骤，也是当前抗疟疾疗法不针对的步骤。 PfCDPK5 最近被确定为一种对寄生虫排出至关重要的激酶。使用最先进的 由于可诱导的蛋白质不稳定系统，现在可以对该必需基因进行功能评估。 PfCDPK5 可诱导敲低的转基因寄生虫在外出前被逮捕。调节 PfCDPK5 水平的能力为研究这种重要激酶的激活机制和下游效应器提供了独特的资源。我们假设 PfCDPK5 是基于钙的出口信号通路的中心介质。该提案的直接目标是获得对 PfCDPK5 激活的分子理解并表征其在出口信号通路中的作用。第一个目标是分子 PfCDPK5 激活的遗传分析将使用转基因寄生虫进行，该寄生虫允许诱导调节必需蛋白质水平。第二个目标是使用候选基因方法和先进的蛋白质组技术来发现 PfCDPK5 的下游底物。这些研究的长期目标和公共卫生影响是确定新的抗疟疾疗法的新靶点。这一长期目标将作为恶性疟原虫中必需的 PfCDPK5 信号通路的分子特征的直接结果而实现。