A UPF0118 family protein with uncharacterized function from the moderate halophile Halobacillus andaensis represents a novel class of Na(+)(Li(+))/H(+) antiporter.
A UPF0118 family protein with uncharacterized function from the moderate halophile Halobacillus andaensis represents a novel class of Na(+)(Li(+))/H(+) antiporter.
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来自中度嗜盐菌安达盐杆菌的 UPF0118 家族蛋白具有未表征的功能，代表了一类新型 Na (Li )/H 逆向转运蛋白
DOI：
10.1038/srep45936
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发表时间：
2017-04-04
影响因子：
4.6
通讯作者：
Jiang J
中科院分区：
文献类型：
--
作者：
Dong P;Wang L;Song N;Yang L;Chen J;Yan M;Chen H;Zhang R;Li J;Abdel-Motaal H;Jiang J
在本研究中，通过在缺乏三个主要Na+/H+反向转运蛋白的大肠杆菌KNabc中的选择，从安达盐芽孢杆菌NEAU-ST 10 - 40 T的基因组DNA中筛选。最终克隆了一个具有Na+（Li+）/H+反向转运活性的基因upf 0118。蛋白质序列比对结果表明，UPF 0118与H. kuroshimensis的Na ~+（Li ~+）/H ~+逆向转运蛋白基因，但与所有已知的特异性Na ~+（Li ~+）/H ~+逆向转运蛋白基因或具有Na ~+（Li ~+）/H ~+逆向转运活性的基因不具有同一性。生长试验、western blot和Na+（Li+）/H+反向转运试验表明，UPF 0118具有pH依赖性Na+（Li+）/H+反向转运活性。系统进化分析表明，UPF 0118与UPF 0118家族的所有同源蛋白在22-82%的同源性范围内聚为一簇，自举值为92%，与已知的Na+（Li+）/H+反向转运蛋白和具有Na+（Li+）/H+反向转运活性的单基因蛋白的同源性较远。综上所述，我们认为UPF 0118应该是一类新的Na+（Li+）/H+反向转运蛋白。据我们所知，这是第一次报告的功能分析的蛋白质与未知的蛋白质功能作为一个代表的UPF 0118家族含有未知功能的结构域，DUF 20。
In this study, genomic DNA was screened from Halobacillus andaensis NEAU-ST10-40T by selection in Escherichia coli KNabc lacking three major Na+/H+ antiporters. One gene designated upf0118 exhibiting Na+(Li+)/H+ antiport activity was finally cloned. Protein alignment showed that UPF0118 shares the highest identity of 81.5% with an unannotated gene encoding a protein with uncharacterized protein function belonging to UPF0118 family from H. kuroshimensis, but shares no identity with all known specific Na+(Li+)/H+ antiporter genes or genes with Na+(Li+)/H+ antiport activity. Growth test, western blot and Na+(Li+)/H+ antiport assay revealed that UPF0118 as a transmembrane protein exhibits pH-dependent Na+(Li+)/H+ antiport activity. Phylogenetic analysis indicated that UPF0118 clustered with all its homologs belonging to UPF0118 family at a wide range of 22–82% identities with the bootstrap value of 92%, which was significantly distant with all known specific single-gene Na+(Li+)/H+ antiporters and single-gene proteins with the Na+(Li+)/H+ antiport activity. Taken together, we propose that UPF0118 should represent a novel class of Na+(Li+)/H+ antiporter. To the best of our knowledge, this is the first report on the functional analysis of a protein with uncharacterized protein function as a representative of UPF0118 family containing the domain of unknown function, DUF20.