Identification of a novel Staphylococcus aureus two-component leukotoxin using cell surface proteomics.
Identification of a novel Staphylococcus aureus two-component leukotoxin using cell surface proteomics.
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DOI：
10.1371/journal.pone.0011634
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发表时间：
2010-07-16
期刊：
影响因子：
3.7
通讯作者：
DeLeo FR
中科院分区：
文献类型：
--
作者：
Ventura CL;Malachowa N;Hammer CH;Nardone GA;Robinson MA;Kobayashi SD;DeLeo FR
金黄色葡萄球菌是一种重要的人类病原体，也是医院和社区细菌感染的主要原因。社区相关耐甲氧西林沙门氏菌金黄色葡萄球菌（CA-MRSA）菌株如USA 300是高毒力的，并且与医院菌株不同，通常在其他健康个体中引起疾病。CA-MRSA增强的毒力部分基于产生高水平分泌分子的能力增加，所述分泌分子促进逃避先天免疫应答。虽然已经取得了进展，但导致CA-MRSA毒力的因素尚未完全确定。我们分析了USA 300菌株LAC的细胞表面蛋白质组（surfome），以更好地了解有助于增强毒力表型的细胞外因子。共113个确定的蛋白质与USA 300在体外生长的后期指数阶段的表面。蛋白A是USA 300最丰富的表面分子，如通过串联质谱法分析肽后的组合Mascot评分所示。出乎意料的是，我们确定了一个以前未表征的双组分白细胞毒素-这里命名为LukS-H和LukF-G（LukGH）-作为两个最丰富的表面相关蛋白的USA 300。兔抗LukG抗体表明USA 300也能将其自由分泌到培养基中。我们使用USA 300的野生型和同基因lukGH缺失菌株结合人PMN孔形成和裂解试验来鉴定该分子为白细胞毒素。此外，LukGH与PVL在体外协同增强人PMNs的溶解，并有助于吞噬后的PMNs的溶解。我们的结论是LukGH是一种新的双组分白细胞毒素，对中性粒细胞具有细胞溶解活性，因此可能有助于S。金黄色葡萄球菌毒力。
Staphylococcus aureus is a prominent human pathogen and leading cause of bacterial infection in hospitals and the community. Community-associated methicillin-resistant S. aureus (CA-MRSA) strains such as USA300 are highly virulent and, unlike hospital strains, often cause disease in otherwise healthy individuals. The enhanced virulence of CA-MRSA is based in part on increased ability to produce high levels of secreted molecules that facilitate evasion of the innate immune response. Although progress has been made, the factors that contribute to CA-MRSA virulence are incompletely defined. We analyzed the cell surface proteome (surfome) of USA300 strain LAC to better understand extracellular factors that contribute to the enhanced virulence phenotype. A total of 113 identified proteins were associated with the surface of USA300 during the late-exponential phase of growth in vitro. Protein A was the most abundant surface molecule of USA300, as indicated by combined Mascot score following analysis of peptides by tandem mass spectrometry. Unexpectedly, we identified a previously uncharacterized two-component leukotoxin–herein named LukS-H and LukF-G (LukGH)-as two of the most abundant surface-associated proteins of USA300. Rabbit antibody specific for LukG indicated it was also freely secreted by USA300 into culture media. We used wild-type and isogenic lukGH deletion strains of USA300 in combination with human PMN pore formation and lysis assays to identify this molecule as a leukotoxin. Moreover, LukGH synergized with PVL to enhance lysis of human PMNs in vitro, and contributed to lysis of PMNs after phagocytosis. We conclude LukGH is a novel two-component leukotoxin with cytolytic activity toward neutrophils, and thus potentially contributes to S. aureus virulence.
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影响因子：
168.9
作者：
DeLeo, Frank R.;Otto, Michael;Kreiswirth, Barry N.;Chambers, Henry F.
通讯作者：
Chambers, Henry F.
影响因子：
3.2
作者：
Jones, Richard C.;Deck, Joanna;Hart, Mark E.
通讯作者：
Hart, Mark E.
影响因子：
3.4
作者：
Gatlin, CL;Pieper, R;Peterson, SN
通讯作者：
Peterson, SN
影响因子：
3.5
作者：
CHOORIT, W;KANEKO, J;KAMIO, Y
通讯作者：
KAMIO, Y
影响因子：
3.7
作者：
Diep, Binh An;Palazzolo-Ballance, Amy M.;Chambers, Henry F.
通讯作者：
Chambers, Henry F.