Identification and molecular characterisation of a gene encoding a member of the insulin receptor family in Echinococcus multilocularis
Identification and molecular characterisation of a gene encoding a member of the insulin receptor family in Echinococcus multilocularis
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DOI：
10.1016/s0020-7519(02)00265-5
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发表时间：
2003-03-01
影响因子：
4
通讯作者：
Brehm, K
中科院分区：
文献类型：
--
作者：
Konrad, C;Kroner, A;Brehm, K
受体激酶在细胞与其环境的沟通中发挥着关键作用，并且可能是宿主细胞因子对内寄生生物影响的重要介质。在本文中我们描述了。首次对寄生蠕虫胰岛素受体家族的受体酪氨酸激酶进行了表征。使用简并性 PCR 方法，我们鉴定并完全表征了多房棘球蚴因子 (EmIR) 的 5.5 kb 编码 DNA，该因子与不同系统发育起源的胰岛素受体表现出显着的同源性。 EmIR 表现出该蛋白质家族的局部结构域，并在相应位置包含所有具有催化作用的重要残基。 EmIR 和已知胰岛素受体之间的一个显着差异是在酪氨酸激酶区域中存在 172 个氨基酸插入片段，但其功能尚不清楚。在酵母双杂交分析中，人胰岛素受体和 EmIR 的配体结合域显示出与人胰岛素相当的亲和力。 EmIR 编码染色体位点 (emir)，如所表征并包含 16.5 kb。 Southern 印迹杂交证明 emir 在多房艾美耳球菌中以单拷贝基因座的形式存在。此外。结构比较表明，埃米尔和哺乳动物和昆虫的胰岛素受体基因源自共同的祖先。基于逆转录酶-聚合酶链反应分析，发现 emir vas 在后头节和原头节这两个幼虫阶段表达。因此，EmIR 是。可能在棘球蚴发育以及与哺乳动物宿主的相互作用中发挥重要作用。 (C) 2003 年澳大利亚寄生虫学协会。由 Elsevier Science Ltd 出版。保留所有权利。
Receptor kinases plan a key role in the communication of cells with their environment and could be important mediators of the effects of host cytokines on endoparasitic organisms. In this paper we describe. for the first time, the characterisation of a receptor tyrosine kinase of the insulin receptor family from a parasitic helminth. Using a degenerative PCR approach, we identified and completely characterised the 5.5 kb coding DNA for an Echinococcus multilocularis factor (EmIR) which displays significant homologies to insulin receptors of different phylogenetic origin. EmIR exhibited a domain structure which is topical for the protein family and contained all catalytically important residues at corresponding positions. One striking difference between EmIR and known insulin receptors was the presence of a 172 amino acid insert in the tyrosine kinase region of, as yet, unknown function. In yeast two-hybrid analyses, the ligand binding domains of the human insulin receptor and of EmIR showed comparable affinity to human insulin. The EmIR encoding chromosomal locus (emir), as characterised and comprised 16.5 kb. Southern blot hybridisations demonstrated that emir is present as a single copy locus in E. multilocularis. Furthermore. structural comparisons indicated that emir and the insulin receptor genes from mammals and insects derive from a common ancestor. Based on reverse transcriptase-polymerase chain reaction analyses, emir vas found to be expressed in the two larval stages metacestode and protoscolex. EmIR is, therefore. likely to play an important role in echinococcal development and possibly also in the interaction with the mammalian host. (C) 2003 Australian Societ for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.