Institute for Microbiology, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
J Proteome Res. 2010 Mar 5;9(3):1579-90. doi: 10.1021/pr901143a.
Most of the Staphylococcus aureus virulence factors are either cell surface exposed or secreted. Here we report a global and quantitative analysis of staphylococcal cell surface-associated proteins using a combination of (14)N(15)N metabolic labeling, biotinylation, and GeLC-MS/MS. To address the important question of S. aureus pathophysiology, we analyzed the influence of the alternative sigma factor sigma(B) on the expression of cell surface-associated proteins. Therefore, we compared the methicillin-resistant S. aureus wild-type strain COL with its sigB mutant, because sigma(B) might play a crucial role in the pattern of the surface proteome. A total of 296 proteins from growing and nongrowing cells could be quantified. One third of these proteins are known as cell surface-associated, including 3 sortase substrates, 3 cell wall-associated proteins, 35 lipo-, 23 membrane-, and 34 signal peptide-containing proteins comparing wild-type and sigB mutant. Fourty nine surface-associated proteins were modulated by sigma(B), including 21 proteins already known to be SigB-dependent or SigB-influenced. More proteins were down- (31 proteins) than up-regulated (18 proteins) in the sigB mutant. Our approach revealed 28 surface-associated proteins not previously reported as SigB-dependent or -influenced, expanding the group of surface-associated proteins and virulence factors modulated by SigB.
大多数金黄色葡萄球菌的毒力因子要么暴露在细胞表面，要么被分泌出来。在这里，我们结合（14）N（15）N 代谢标记、生物素化和 GeLC-MS/MS 对金黄色葡萄球菌细胞表面相关蛋白进行了全面和定量分析。为了解决金黄色葡萄球菌病理生理学的重要问题，我们分析了替代 sigma 因子 sigma(B) 对细胞表面相关蛋白表达的影响。因此，我们比较了耐甲氧西林金黄色葡萄球菌野生型菌株 COL 与其 sigB 突变体，因为 sigma(B) 可能在表面蛋白质组模式中发挥关键作用。总共可以定量 296 种来自生长和非生长细胞的蛋白质。其中三分之一是已知的细胞表面相关蛋白，包括 3 种分选酶底物、3 种细胞壁相关蛋白、35 种脂类、23 种膜和 34 种信号肽含蛋白，与野生型和 sigB 突变体相比。sigma(B) 调节了 49 种表面相关蛋白，包括 21 种已知依赖 SigB 或受 SigB 影响的蛋白。在 sigB 突变体中，下调的蛋白（31 个）多于上调的蛋白（18 个）。我们的方法揭示了 28 种以前未报道为 SigB 依赖或受 SigB 影响的表面相关蛋白，扩大了受 SigB 调节的表面相关蛋白和毒力因子的群体。
