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曼氏血吸虫(吸虫纲:复殖目)中一个与Shaker相关的电压门控钾通道基因的克隆及功能表达

Cloning and functional expression of a Shaker-related voltage-gated potassium channel gene from Schistosoma mansoni (Trematoda: Digenea).

作者信息

Kim E, Day T A, Bennett J L, Pax R A

机构信息

Department of Pharmacology and Toxicology, Michigan State University, East Lansing 48824.

出版信息

Parasitology. 1995 Feb;110 ( Pt 2):171-80. doi: 10.1017/s0031182000063939.

Abstract

We have isolated a cDNA (SKv1.1) encoding a Shaker-related K+ channel from an adult cDNA library of the human parasitic trematode Schistosoma mansoni. The deduced amino acid sequence (512 aa, 56.5 kDa) contains 6 putative membrane-spanning domains (S1-S6) and a pore-forming domain (H5). SKv1.1 is grouped in the Shaker family, but forms a unique branch within this family, on the basis of dendrogram analysis. SKv1.1 shows significant sequence identity with most other Shaker channels, with 64-74% identity in the core region (S1-S6). It has the highest sequence identity with the K+ channel (Ak01a) from Aplysia. Northern blot analysis detected a single primary transcript of 2.8 kb. Southern blot analysis indicated that SKv1.1 is present as a single copy in the genomic DNA of S. mansoni. Expression of SKv1.1 in Xenopus oocytes produced a rapidly activating and inactivating outward K+ current which is highly sensitive to 4-aminopyridine, but is insensitive to tetraethylammonium, mast cell degranulating peptide, dendrotoxin and charybdotoxin. The presence of a Shaker homologue in Schistosoma suggests that Sh subfamilies may exist in other lower invertebrates as well as platyhelminths.

摘要

我们从人类寄生吸虫曼氏血吸虫的成体cDNA文库中分离出了一个编码与Shaker相关的钾离子通道的cDNA(SKv1.1)。推导的氨基酸序列(512个氨基酸,56.5 kDa)包含6个假定的跨膜结构域(S1 - S6)和一个孔形成结构域(H5)。基于系统树分析,SKv1.1归属于Shaker家族,但在该家族中形成一个独特的分支。SKv1.1与大多数其他Shaker通道显示出显著的序列同一性,在核心区域(S1 - S6)具有64 - 74%的同一性。它与海兔的钾离子通道(Ak01a)具有最高的序列同一性。Northern印迹分析检测到一个2.8 kb的单一初级转录本。Southern印迹分析表明SKv1.1在曼氏血吸虫的基因组DNA中以单拷贝形式存在。SKv1.1在非洲爪蟾卵母细胞中的表达产生了一种快速激活和失活的外向钾电流,该电流对4 - 氨基吡啶高度敏感,但对四乙铵、肥大细胞脱颗粒肽、树突毒素和蝎毒素不敏感。血吸虫中存在Shaker同源物表明Sh亚家族可能也存在于其他低等无脊椎动物以及扁形动物中。

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