Differential Membrane Proteome Analysis Reveals Novel Proteins Involved in the Degradation of Aromatic Compounds in Geobacter metallireducens
Differential Membrane Proteome Analysis Reveals Novel Proteins Involved in the Degradation of Aromatic Compounds in Geobacter metallireducens
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DOI：
10.1074/mcp.m900061-mcp200
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发表时间：
2009-09-01
影响因子：
7
通讯作者：
Boll, Matthias
中科院分区：
文献类型：
--
作者：
Heintz, Dimitri;Gallien, Sebastien;Boll, Matthias
芳香族化合物包括一大类天然和人造化合物，其中许多对环境和人类健康有相当大的影响。在芳香族化合物降解厌氧细菌中，芳香族分解代谢的中心中间体苯甲酰辅酶A受到去芳香化苯甲酰辅酶A还原酶（bcr）的攻击。兼性厌氧菌中存在atp依赖性BCR。相比之下，先前对专性厌氧模式生物金属还原Geobacter metallireducens的可溶性蛋白质组的分析发现了编码完全不同的去芳香化BCR的基因。预计相应的BamBCDEFGHI配合物包含可溶性钼或钨、硒代半胱氨酸和含FeS簇的组分。为了阐明专性厌氧细菌降解芳香族化合物的关键过程，采用基于质谱的光谱计数方法测定了生长在苯甲酸酯和醋酸酯上的G. metallireducens膜蛋白丰度的差异。采用一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳-液相色谱-串联质谱联用技术，共鉴定出931种蛋白质。新发现了几种参与芳香化合物降解的膜相关蛋白，包括与含NiFe/血红素b和能量转换氢化酶、细胞色素bd氧化酶、异化硝酸盐还原酶和钨酸盐atp结合盒转运系统模块相似的蛋白。采用定量逆转录pcr分析差异表达基因的转录调控；此外，还测定了苯甲酸酯诱导的氢化酶和硝酸还原酶的体外活性。获得的结果为专性厌氧细菌中芳香族化合物的降解提供了新的见解。中国生物医学工程学报，2009；
Aromatic compounds comprise a large class of natural and man-made compounds, many of which are of considerable concern for the environment and human health. In aromatic compound-degrading anaerobic bacteria the central intermediate of aromatic catabolism, benzoyl coenzyme A, is attacked by dearomatizing benzoyl-CoA reductases (BCRs). An ATP-dependent BCR has been characterized in facultative anaerobes. In contrast, a previous analysis of the soluble proteome from the obligately anaerobic model organism Geobacter metallireducens identified genes putatively coding for a completely different dearomatizing BCR. The corresponding BamBCDEFGHI complex is predicted to comprise soluble molybdenum or tungsten, selenocysteine, and FeS cluster-containing components. To elucidate key processes involved in the degradation of aromatic compounds in obligately anaerobic bacteria, differential membrane protein abundance levels from G. metallireducens grown on benzoate and acetate were determined by the MS-based spectral counting approach. A total of 931 proteins were identified by combining one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis with liquid chromatography-tandem mass spectrometry. Several membrane-associated proteins involved in the degradation of aromatic compounds were newly identified including proteins with similarities to modules of NiFe/heme b-containing and energy-converting hydrogenases, cytochrome bd oxidases, dissimilatory nitrate reductases, and a tungstate ATP-binding cassette transporter system. The transcriptional regulation of differentially expressed genes was analyzed by quantitative reverse transcription-PCR; in addition benzoate-induced in vitro activities of hydrogenase and nitrate reductase were determined. The results obtained provide novel insights into the poorly understood degradation of aromatic compounds in obligately anaerobic bacteria. Molecular & Cellular Proteomics 8: 2159-2169, 2009.