ASPARAGINYL TRNA SYNTHETASE OF BRUGIA MALAYI

氨基酰基-tRNA合成酶(AARS)是一类重要的 在蛋白质生物合成中起基础作用的酶 线虫。这项建议的总体目标是理解 中和抗体介导的氨酰化现象 重组马来丝虫天冬酰胺tRNA合成酶的抑制作用 (ASNRS)。这项工作将主要集中在识别和 淋巴丝虫病患者亚群的特征 产生中和抗体的流行区，以及 中和抗体的免疫学性质。这项拨款建议 三个具体目标。首先，一个两步筛选过程将确定 患有自发性精神分裂症的人患上马来亚巴氏杆菌AsnRS：tRNA (Asn)从可溶性HeLa细胞提取物中提取出来的复合体。第二，分歧 或中和识别的表位与非中和识别的表位的相似性 中和抗体将通过一组蛋白质印迹分析进行研究。 重叠的合成肽，对应于完整的马来芽胞杆菌 AsnRS.第三，体外氨基酰化试验的特异性 将通过过量生产和重组人的纯化来改善 秀丽线虫tRNA(ASN)基因。从长远来看， 从这些研究中获得的见解将提高我们对 丝虫中AARS家族的基因，有助于了解物种的基础- AsnRS中和抗体的特异性。拟议的研究还包括 将表明自身抗体和 丝虫AsnRs。这些目标将同时提供一个 认为(1)寄生虫AsnRs是一种极好的 合理的抗丝虫病药物设计的目标，以及(2)潜力 寄生虫AsnRs在诱导小鼠自身免疫现象中的作用 淋巴丝虫病。

Aminoacyl-tRNA synthetases (AARS) are an important family of enzymes which play a fundamental role in protein biosynthesis in nematodes. The broad objective of this proposal is to understand the phenomenon of neutralizing antibody mediated aminoacylation inhibition of a recombinant Brugia malayi asparaginyl-tRNA synthetase (AsnRS). This work will focus primarly on identification and characterization of the subset of persons from lymphatic filariasis endemic areas who develop neutralizing antibodies, and the immunological properties of neutralizing antibody. This grant proposes three specific aims. First, a two step screening process will identify persons who have spontaneoulsy developed B. Malayi AsnRS:tRNA (Asn) complex from soluble HeLa cell extracts. Secondly, differences or similarities between epitopes recognized by neutralizing and non- neutralizing antibodies will be studied by western blot analysis of a set of overlapping synthetic peptides, corresponding to the full B. malayi AsnRS. Thirdly, the specificity of an in vitro aminoacylation assay will be improved by overproduction and purification of a recombinant tRNA (Asn) gene derived from Caenorhabditis elegans. Long-term, insight from these studies will improve both our general understanding of AARS family in filaria and help to understand the basis for species- specificity of AsnRS neutralizing antibody. The proposed studies also will indicate the basis for cross reactivity between autoantibody and the filarial AsnRS. These goals will simultaneously provide a foundation on which to consider (1) the parasite AsnRS as an excellent target for rational anti-filarial drug design, and (2) the potential role of parasite AsnRS in the induction of autoimmune phenomenon in lymphatic filariasis.