Almárcegui Rodrigo J, Navarro Claudio A, Paradela Alberto, Albar Juan Pablo, von Bernath Diego, Jerez Carlos A
Laboratory of Molecular Microbiology and Biotechnology, Department of Biology, Faculty of Sciences, University of Chile, Santiago, Chile.
Proteomics Laboratory, National Biotechnology Center, Madrid, Spain.
Res Microbiol. 2014 Nov;165(9):761-72. doi: 10.1016/j.resmic.2014.07.005. Epub 2014 Jul 18.
The response of Acidithiobacillus ferrooxidans ATCC 23270 to copper was analyzed in sulfur-grown cells by using quantitative proteomics. Forty-seven proteins showed altered levels in cells grown in the presence of 50 mM copper sulfate. Of these proteins, 24 were up-regulated and 23 down-regulated. As seen before in ferrous iron-grown cells, there was a notorious up-regulation of RND-type Cus systems and different RND-type efflux pumps, indicating that these proteins are very important in copper resistance. Copper also triggered the down-regulation of the major outer membrane porin of A. ferrooxidans in sulfur-grown bacteria, suggesting they respond to the metal by decreasing the influx of cations into the cell. On the contrary, copper in sulfur-grown cells caused an overexpression of putative TadA and TadB proteins known to be essential for biofilm formation in bacteria. Surprisingly, sulfur-grown microorganisms showed increased levels of proteins related with energy generation (rus and petII operons) in the presence of copper. Although rus operon is overexpressed mainly in cells grown in ferrous iron, the up-regulation of rusticyanin in sulfur indicates a possible role for this protein in copper resistance as well. Finally, copper response in A. ferrooxidans appears to be influenced by the substrate being oxidized by the microorganism.
利用定量蛋白质组学分析了嗜酸氧化亚铁硫杆菌ATCC 23270在硫培养基中生长的细胞对铜的响应。在添加50 mM硫酸铜的培养基中生长的细胞中,有47种蛋白质的水平发生了变化。其中,24种蛋白质上调,23种蛋白质下调。正如之前在亚铁培养基中生长的细胞中所观察到的,RND型Cus系统和不同的RND型外排泵显著上调,这表明这些蛋白质在抗铜方面非常重要。铜还导致硫培养基中生长的嗜酸氧化亚铁硫杆菌的主要外膜孔蛋白下调,这表明它们通过减少阳离子流入细胞来响应金属。相反,硫培养基中生长的细胞中的铜导致已知对细菌生物膜形成至关重要的假定TadA和TadB蛋白过表达。令人惊讶的是,在有铜存在的情况下,硫培养基中生长的微生物中与能量产生相关的蛋白质(rus和petII操纵子)水平升高。虽然rus操纵子主要在亚铁培养基中生长的细胞中过表达,但硫培养基中rusticyanin的上调表明该蛋白在抗铜方面也可能发挥作用。最后,嗜酸氧化亚铁硫杆菌对铜的响应似乎受微生物氧化的底物影响。