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TbIRK 是一种来自布氏锥虫的无特征序列钾通道,定位于液泡型酸性磷酸酶体。

TbIRK is a signature sequence free potassium channel from Trypanosoma brucei locating to acidocalcisomes.

机构信息

Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.

Swiss Tropical and Public Health Institute, Basel, Switzerland.

出版信息

Sci Rep. 2017 Apr 6;7(1):656. doi: 10.1038/s41598-017-00752-1.

Abstract

Potassium channels from prokaryotes and eukaryotes are usually recognized by a typical amino acid sequence TXTGY(F)G representing the ionic selectivity filter. Using a screening approach with ion channel family profiles but without the above motif, we identified a gene in Trypanosoma brucei that exhibits homology to inward rectifying potassium channels. We report here cloning of this ion channel named TbIRK. The protein is localized to acidocalcisomes in procyclic and in bloodstream form parasites. Functional properties of this channel were established after expression in Xenopus oocytes. Currents recorded in potassium medium show inward rectification and little time dependence. Surprisingly, this channel retains selectivity for potassium ions over sodium ions >7, in spite of the lack of the classical selectivity filter. The sequence GGYVG was predicted in silico to replace this filter motif. Point mutations of the corresponding glycine residues confirmed this at the functional level. The channel is inhibited by caesium ions but remains unaffected by barium ions up to 10 mM. TbIRK is to our knowledge the first potassium channel in T. brucei that localizes to the acidocalcisomes, organelles involved in the storage of phosphates and the response to osmotic stress that occurs during the life cycle of trypanosomes.

摘要

原核生物和真核生物的钾通道通常通过代表离子选择性过滤器的典型氨基酸序列 TXTGY(F)G 来识别。我们使用带有离子通道家族图谱的筛选方法,但没有上述模体,鉴定出在布氏锥虫中存在一个与内向整流钾通道具有同源性的基因。我们在此报道了该离子通道的克隆,命名为 TbIRK。该蛋白定位于前鞭毛体和血液阶段寄生虫的酸性囊泡中。在非洲爪蟾卵母细胞中表达后,建立了该通道的功能特性。在钾介质中记录的电流表现出内向整流和几乎没有时间依赖性。令人惊讶的是,尽管缺乏经典的选择性过滤器,但该通道仍对钾离子的选择性高于钠离子 >7。通过计算机预测序列 GGYVG 来替代该过滤器模体。相应甘氨酸残基的点突变在功能水平上证实了这一点。该通道被铯离子抑制,但在 10mM 以下不受钡离子的影响。据我们所知,TbIRK 是布氏锥虫中第一个定位于酸性囊泡的钾通道,酸性囊泡参与磷酸盐的储存以及在锥虫生命周期中发生的渗透压应激反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6a3f/5429665/03570a9f6e9c/41598_2017_752_Fig1_HTML.jpg

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