ASPARAGINYL TRNA SYNTHETASE OF BRUGIA MALAYI

氨酰-tRNA合成酶（阿尔斯）是一个重要的 在蛋白质生物合成中起重要作用的酶， 线虫 本提案的总体目标是了解 中和抗体介导的氨酰化现象 重组马来丝虫天冬酰胺酰-tRNA合成酶抑制 （AsnRS）。 这项工作将主要侧重于查明和 淋巴丝虫病患者亚群的特征 流行地区的人产生中和抗体， 中和抗体的免疫学特性。该基金建议 三个具体目标。 首先，两步筛选过程将确定 自发发展出B的人。马来语AsnRS：tRNA (Asn)可溶性HeLa细胞提取物的复合物。 第二，差异 或通过中和和非中和识别的表位之间的相似性， 中和抗体将通过一组免疫印迹分析来研究 重叠的合成肽，对应于完整的B。马来丝虫 AsnRS。 第三，体外氨酰化试验的特异性 将通过过量生产和纯化重组体 来源于秀丽隐杆线虫的tRNA（Asn）基因。 长期来看， 从这些研究中获得的见解将提高我们对 阿尔斯家族在丝虫中的作用，并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 将表明自身抗体与 丝虫AsnRS 这些目标将同时提供一个 在此基础上考虑（1）寄生虫AsnRS作为一个优秀的 合理抗丝虫药物设计的目标，和（2）潜在的 寄生虫AsnRS在诱导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.