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从海洋盐单胞菌中鉴定一种新型的两成分 Na(Li, K)/H 反向转运蛋白。

Characterization of a novel two-component Na(Li, K)/H antiporter from Halomonas zhaodongensis.

机构信息

Department of Microbiology and Biotechnology, College of Life Sciences, Northeast Agricultural University, Harbin, China.

出版信息

Sci Rep. 2017 Jun 26;7(1):4221. doi: 10.1038/s41598-017-04236-0.

DOI:10.1038/s41598-017-04236-0
PMID:28652569
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5484666/
Abstract

In this study, genomic DNA was screened for novel Na/H antiporter genes from Halomonas zhaodongensis by selection in Escherichia coli KNabc lacking three major Na/H antiporters. Co-expression of two genes designated umpAB, encoding paired homologous unknown membrane proteins belonging to DUF1538 (domain of unknown function with No. 1538) family, were found to confer E. coli KNabc the tolerance to 0.4 M NaCl and 30 mM LiCl, and an alkaline pH resistance at 8.0. Western blot and co-immunoprecipitation establish that UmpAB localize as a hetero-dimer in the cytoplasmic membranes. Functional analysis reveals that UmpAB exhibit pH-dependent Na(Li, K)/H antiport activity at a wide pH range of 6.5 to 9.5 with an optimal pH at 9.0. Neither UmpA nor UmpB showed homology with known single-gene or multi-gene Na/H antiporters, or such proteins as ChaA, MdfA, TetA(L), Nap and PsmrAB with Na/H antiport activity. Phylogenetic analysis confirms that UmpAB should belong to DUF1538 family, which are significantly distant with the above-mentioned proteins with Na/H antiport activity. Taken together, we propose that UmpAB represent a novel two-component Na(Li, K)/H antiporter. To the best of our knowledge, this is the first report on the functional analysis of unknown membrane proteins belonging to DUF1538 family.

摘要

在这项研究中，通过在缺乏三种主要 Na/H 反向转运蛋白的大肠杆菌 KNabc 中的选择，从海洋盐单胞菌中筛选新型 Na/H 反向转运蛋白基因的基因组 DNA。共表达两个基因命名为 umpAB，它们编码属于 DUF1538（功能未知域，编号 1538）家族的成对同源未知膜蛋白，发现它们使大肠杆菌 KNabc 能够耐受 0.4 M NaCl 和 30 mM LiCl，并在 pH8.0 时具有碱性 pH 抗性。Western blot 和共免疫沉淀证实 UmpAB 定位于细胞质膜中作为异源二聚体。功能分析表明，UmpAB 在 pH6.5 至 9.5 的宽 pH 范围内表现出 pH 依赖性的 Na(Li，K)/H 反向转运活性，最佳 pH 值为 9.0。UmpA 和 UmpB 均与已知的单基因或多基因 Na/H 反向转运蛋白或具有 Na/H 反向转运活性的 ChaA、MdfA、TetA(L)、Nap 和 PsmrAB 等蛋白质没有同源性。系统发育分析证实 UmpAB 应属于 DUF1538 家族，与具有 Na/H 反向转运活性的上述蛋白质明显不同。综上所述，我们提出 UmpAB 代表一种新型的双组分 Na(Li，K)/H 反向转运蛋白。据我们所知，这是首次对属于 DUF1538 家族的未知膜蛋白的功能分析进行报道。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/ff9aa83a8c75/41598_2017_4236_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/eb3bbaa719c9/41598_2017_4236_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/7e0e09984286/41598_2017_4236_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/5f77fd0ab8d4/41598_2017_4236_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/6c8b0c59d036/41598_2017_4236_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/1a87252f7796/41598_2017_4236_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/6c262246275c/41598_2017_4236_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/2e3cd4bce8dc/41598_2017_4236_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/b3312973a66f/41598_2017_4236_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/ff9aa83a8c75/41598_2017_4236_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/eb3bbaa719c9/41598_2017_4236_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/7e0e09984286/41598_2017_4236_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/5f77fd0ab8d4/41598_2017_4236_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/6c8b0c59d036/41598_2017_4236_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/1a87252f7796/41598_2017_4236_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/6c262246275c/41598_2017_4236_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/2e3cd4bce8dc/41598_2017_4236_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/b3312973a66f/41598_2017_4236_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d10b/5484666/ff9aa83a8c75/41598_2017_4236_Fig9_HTML.jpg

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