A Comparative Quantitative Proteomic Study Identifies New Proteins Relevant for Sulfur Oxidation in the Purple Sulfur Bacterium Allochromatium vinosum
A Comparative Quantitative Proteomic Study Identifies New Proteins Relevant for Sulfur Oxidation in the Purple Sulfur Bacterium Allochromatium vinosum
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DOI：
10.1128/aem.04182-13
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发表时间：
2014-01
影响因子：
4.4
通讯作者：
T. Weißgerber;M. Sylvester;Lena Kröninger;C. Dahl
中科院分区：
文献类型：
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作者：
T. Weißgerber;M. Sylvester;Lena Kröninger;C. Dahl
摘要 在本研究中，我们比较了 Allochromatium vinosum 在硫化物、硫代硫酸盐和元素硫存在下光合自养生长时的蛋白质组响应与该生物体在苹果酸上光合异养生长时的蛋白质组响应。应用串联质量标签分析以及二维 (2D) PAGE，我们通过鉴定至少两个肽 (59.2%) 检测到了 3,302 种预测蛋白质中的 1,955 种，并对 1,848 种已鉴定蛋白质进行了定量。观察到 385 种蛋白质的相对蛋白质含量发生改变（≥1.5 倍），相当于量化的 A. vinosum 蛋白质组的 20.8%。在还原硫化合物存在的情况下，大量表现出相对蛋白质水平强烈增强的蛋白质是氧化硫代谢过程中的重要参与者，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，并允许鉴定参与氧化硫代谢的新基因/蛋白质。选择在转录组和蛋白质组水平上均表现出强烈反应的一个基因簇（hyd；Alvin_2036-Alvin_2040）和一种假设蛋白（Alvin_2107）用于各自突变株的基因失活和表型分析，这验证了所谓的Isp氢化酶超级复合物对于有效氧化硫化物的重要性以及Alvin_2107对于硫中储存的硫的氧化的关键作用球状变为亚硫酸盐。此外，我们分析了硫球蛋白质组并鉴定了一种新的硫球蛋白质（SgpD；Alvin_2515）。
ABSTRACT 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).