Weissgerber Thomas, Sylvester Marc, Kröninger Lena, Dahl Christiane
Institut für Mikrobiologie & Biotechnologie, Rheinische Friedrich-Wilhelms-Universität Bonn, Bonn, Germany.
Appl Environ Microbiol. 2014 Apr;80(7):2279-92. doi: 10.1128/AEM.04182-13. Epub 2014 Jan 31.
In the present study, we compared the proteome response of Allochromatium vinosum when growing photoautotrophically in the presence of sulfide, thiosulfate, and elemental sulfur with the proteome response when the organism was growing photoheterotrophically on malate. Applying tandem mass tag analysis as well as two-dimensional (2D) PAGE, we detected 1,955 of the 3,302 predicted proteins by identification of at least two peptides (59.2%) and quantified 1,848 of the identified proteins. Altered relative protein amounts (≥1.5-fold) were observed for 385 proteins, corresponding to 20.8% of the quantified A. vinosum proteome. A significant number of the proteins exhibiting strongly enhanced relative protein levels in the presence of reduced sulfur compounds are well documented essential players during oxidative sulfur metabolism, e.g., the dissimilatory sulfite reductase DsrAB. Changes in protein levels generally matched those observed for the respective relative mRNA levels in a previous study and allowed identification of new genes/proteins participating in oxidative sulfur metabolism. One gene cluster (hyd; Alvin_2036-Alvin_2040) and one hypothetical protein (Alvin_2107) exhibiting strong responses on both the transcriptome and proteome levels were chosen for gene inactivation and phenotypic analyses of the respective mutant strains, which verified the importance of the so-called Isp hydrogenase supercomplex for efficient oxidation of sulfide and a crucial role of Alvin_2107 for the oxidation of sulfur stored in sulfur globules to sulfite. In addition, we analyzed the sulfur globule proteome and identified a new sulfur globule protein (SgpD; Alvin_2515).
在本研究中,我们比较了嗜硫红假单胞菌(Allochromatium vinosum)在硫化物、硫代硫酸盐和元素硫存在下进行光合自养生长时的蛋白质组反应,以及该生物体在苹果酸上进行光合异养生长时的蛋白质组反应。应用串联质量标签分析以及二维(2D)聚丙烯酰胺凝胶电泳,我们通过鉴定至少两个肽段,在3302个预测蛋白中检测到了1955个(59.2%),并对其中1848个已鉴定蛋白进行了定量。观察到385个蛋白的相对蛋白量发生了改变(≥1.5倍),这相当于定量的嗜硫红假单胞菌蛋白质组的20.8%。在还原态硫化合物存在下,大量相对蛋白水平显著提高的蛋白是氧化硫代谢过程中已充分记录的关键参与者,例如异化亚硫酸盐还原酶DsrAB。蛋白质水平的变化通常与先前研究中观察到的相应相对mRNA水平的变化相匹配,并有助于鉴定参与氧化硫代谢的新基因/蛋白。选择了一个在转录组和蛋白质组水平上均表现出强烈反应的基因簇(hyd;Alvin_2036 - Alvin_2040)和一个假定蛋白(Alvin_2107),对各自的突变菌株进行基因失活和表型分析,这证实了所谓的Isp氢化酶超复合物对硫化物高效氧化的重要性,以及Alvin_2107对硫球中储存的硫氧化为亚硫酸盐的关键作用。此外,我们分析了硫球蛋白质组,并鉴定出一种新的硫球蛋白(SgpD;Alvin_2515)。