A new family of secreted adhesive proteins in Plasmodium

疟原虫中分泌的粘附蛋白的一个新家族

• 批准号: 6847405
• 负责人: THOMAS J TEMPLETON
• 金额: $ 33.6万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6847405
• 关键词: Anopheles; Apicomplexa; Plasmodium; Plasmodium berghei; Plasmodium falciparum; antiserum; biochemical evolution; clinical research; disease /disorder model; disease vectors; functional /structural genomics; host organism interaction; human tissue; immunoelectron microscopy; laboratory mouse; laboratory rabbit; laboratory rat; life cycle; malaria; microorganism culture; parasite infection mechanism; protein localization; protein quantitation /detection; protein structure; proteomics; protozoal genetics

DESCRIPTION (provided by the applicant): Despite decades of pharmacological and pesticide intervention, Plasmodium remains the most prominent pathogenic protozoan, responsible for widespread morbidity and mortality in tropical regions of the world. Progress in the "post-genomic" era, utilizing whole genome techniques such as micro-arrays, proteomics and genome sequence annotation, is rapidly providing research targets, but ultimate success will necessitate a gene-by-gene pursuit of biological questions concerning newly identified drug and vaccine candidates. Via whole genome sequence analysis of Plasmodium and Cryptosporidium we are determining the provenance of extracellular domains likely arising in the parasitic adaptation of the pathogenic Apicomplexa. A select set of genes identified in this analysis share the presence of an LCCL domain and have striking multi-domain architectures composed of animal- and bacterial-derived extracellular adhesive modules. These genes are singularly remarkable among apicomplexan multi-domain extracellular proteins in that orthologs are conserved across the apicomplexan clade and thus these proteins are likely to serve a conserved biological role widespread in the pathogenic Apicomplexa. Transcript and protein expression studies indicate that the Plasmodium orthologs, herein termed PCCp1, PCCp2 and PCCp3, are expressed in gametocyte stages and during gametogenesis, and thus they are promising candidates for study of biological function during mosquito transmission. Preliminary protein localization studies in mature gametocytes indicate that PCCp proteins are trafficked to the parasite surface or a sub-membranous compartment prior to secretion during gametogenesis. It is a goal of this proposal to refine cellular localization via immunoelectron microscopy. As a means to investigate biological function, PCCp1, PCCp2 and PCCp3 genes will be disrupted in P. falciparum cultured parasites and in the rodent malaria model, P. berghei. An assessment of phenotype will include intraerythrocytic stages as well as investigation of a role in transmission to anopheline mosquitoes. Preliminary results with an antiserum recognizing PfCCp1 LCCL domains indicate transmission-blocking activity in mosquito membrane feeds and thus a component of this proposal is to determine the candidacy of the PCCp proteins in a multi-subunit transmission blocking vaccine (TBV).

描述（由申请人提供）：尽管有数十年的药理学和农药干预，疟原虫仍然是最突出的致病原生动物，负责世界热带地区的广泛发病率和死亡率。在“基因组后”时代的进步，利用整个基因组技术，例如微阵列，蛋白质组学和基因组序列注释，迅速提供了研究目标，但最终的成功将需要对生物学问题进行基因追求，从而使新鉴定的新鉴定的药物和疫苗候选者对生物学问题进行基因追求。通过疟原虫和隐孢子虫的整个基因组序列分析，我们确定了在致病性apicomplexa的寄生虫适应中可能产生的细胞外域的来源。在此分析中鉴定出的一组基因共享LCCL结构域的存在，并具有由动物和细菌衍生的细胞外粘合剂模块组成的引人注目的多域结构。这些基因在Apicomplexan多域外细胞外蛋白中是奇异的，因为直系同源物在整个Apicomplexan进化枝之间是保守的，因此这些蛋白质可能在致病性apicomplexa中具有保守的生物学作用。转录本和蛋白质表达研究表明，本文称为PCCP1，PCCP2和PCCP3的疟原虫在配子细胞阶段和配子发生过程中表达，因此它们是在蚊子传播过程中研究生物学功能的有前途的候选者。在成熟的配子细胞中的初步蛋白质定位研究表明，在配子发生过程中分泌之前，PCCP蛋白在分泌之前将PCCP蛋白运用到寄生虫表面或下膜下室。该建议是通过免疫电子显微镜来完善细胞定位的目标。作为研究生物学功能的一种手段，PCCP1，PCCP2和PCCP3基因将在恶性疟原虫培养的寄生虫和啮齿动物疟疾模型P. berghei中破坏。对表型的评估将包括肠内阶段以及研究传播到蚊子蚊子中的作用。鉴定PFCCP1 LCCL结构域的抗血清的初步结果表明蚊子膜进料中的传输阻滞活性，因此该提案的一个组成部分是确定多支化合物传输阻断疫苗（TBV）中PCCP蛋白的鉴定。