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基于 iTRAQ 的费氏弧菌 BAP-1 诱导的荧蒽差异表达蛋白的比较蛋白质组学分析。

iTRAQ-based comparative proteomic analysis of differentially expressed proteins in Rhodococcus sp. BAP-1 induced by fluoranthene.

机构信息

College of Water Sciences, Beijing Normal University, 100875 Beijing, China.

College of Environment and Resource, Guangxi Normal University, 541004 Guilin, Guangxi, China.

出版信息

Ecotoxicol Environ Saf. 2019 Mar;169:282-291. doi: 10.1016/j.ecoenv.2018.11.022. Epub 2018 Nov 17.

Abstract

To reveal the molecular mechanism at the level of regulation of proteins in Rhodococcus sp. BAP-1 induced by fluoranthene comparative proteomic analysis was performed on proteins extracted from fluoranthene-exposed cells on 1 d, 3 d, 6 d and 8 d compared with control cells using isobaric tags for relative and absolute quantization (iTRAQ) labeling and LC-MS/MS analysis to access differentially expressed proteins. As a result, we detected a total of 897 significantly differentially expressed proteins, including 30 shared proteins in four comparison clusters. We were able to short-list 190, 329, 101 and 90 proteins that were over-represented, and 394, 234, 65 and 49 under-represented proteins, in 1d/control, 3d/control, 6d/control and 8d/control comparisons, respectively. Functional analysis relied on Clusters of Orthologous Groups (COG), gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that fluoranthene significantly altered the expression of proteins involved in metabolic and biosynthesis processes. Furthermore, BAP-1 up-regulates aldehyde dehydrogenase, cytochrome c oxidase, and oligopeptide transport ATP-binding protein, while down-regulates several other proteins in order to adapt to fluoranthene exposure. These findings provide important clues to reveal fluoranthene degradation mechanism in BAP-1 and promote its bioremediation applications.

摘要

为了揭示荧光蒽诱导的罗德里格斯氏菌 BAP-1 中蛋白质调控水平的分子机制,我们采用同位素相对标记与绝对定量(iTRAQ)标记和 LC-MS/MS 分析,对暴露于荧光蒽的细胞和对照组细胞在第 1、3、6 和 8 天提取的蛋白质进行了比较蛋白质组学分析。结果,我们共检测到 897 个差异表达的蛋白质,其中 4 个比较聚类中共有 30 个蛋白质。我们能够将 1d/control、3d/control、6d/control 和 8d/control 比较中分别过表达的 190、329、101 和 90 个蛋白,以及分别低表达的 394、234、65 和 49 个蛋白进行了筛选。功能分析依赖于同源基因簇(COG)、基因本体论(GO)和京都基因与基因组百科全书(KEGG),结果表明荧光蒽显著改变了参与代谢和生物合成过程的蛋白质的表达。此外,BAP-1 上调了醛脱氢酶、细胞色素 c 氧化酶和寡肽转运 ATP 结合蛋白,而下调了其他几种蛋白质,以适应荧光蒽的暴露。这些发现为揭示 BAP-1 中荧光蒽降解机制提供了重要线索,并促进了其生物修复应用。

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