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首页> 外文期刊>Biochimica et biophysica acta. Biomembranes >Surface proteins and the formation of biofilms by Staphylococcus aureus
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Surface proteins and the formation of biofilms by Staphylococcus aureus

机译：表面蛋白质和金黄色葡萄球菌的形成生物膜

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Staphylococcus aureus biofilms pose a serious clinical threat as reservoirs for persistent infections. Despite this clinical significance, the composition and mechanism of formation of S. aureus biofilms are unknown. To address these problems, we used solid-state NMR to examine S. aureus (SA113), a strong biofilm-forming strain. We labeled whole cells and cell walls of planktonic cells, young bitifilms formed for 12-24 h after stationary phase, and more mature biofilms formed for up to 60 h after stationary phase. All samples were labeled either by (i) [N-15]glycine and L-[1-C-13] threonine, or in separate experiments, by (ii) L-[2-C-13,N-15]leucine. We then measured C-13-N-15 direct bonds by C{N} rotational-echo double resonance (REDOR). The increase in peptidoglycan stems that have bridges connected to a surface protein was determined directly by a cell-wall double difference (biofilm REDOR difference minus planktonic REDOR difference). This procedure eliminates errors arising from differences in N-15 isotopic enrichments and from the routing of C-13 label from threonine degradation to glycine. For both planktonic cells and the mature biofilm, 20% of pentaglycyl bridges are not cross-linked and are potential surface-protein attachment sites. None of these sites has a surface protein attached in the planktonic cells, but one-fourth have a surface protein attached in the mature biofilm. Moreover, the leucine-label shows that the concentration of beta-strands in leucine-rich regions doubles in the mature biofilm. Thus, a primary event in establishing a S. aureus biofilm is extensive decoration of the cell surface with surface proteins that are linked covalently to the cell wall and promote cell-cell adhesion.
机译：金黄色葡萄球菌生物膜构成严重的临床威胁作为持续感染的储层。尽管这种临床意义，但形成了金黄色葡萄球菌的形成和机制。为了解决这些问题，我们使用固态NMR来检查金黄色葡萄球菌（SA113），一种强烈的生物膜形成菌株。我们标记了浮游细胞的整个细胞和细胞壁，在固定相后形成12-24小时的幼虫，并且在固定相后更具成熟的生物膜形成最多60小时。所有样品通过（I）[N-15]甘氨酸和L-[1-C-13]苏氨酸或单独的实验标记，通过（II）L- [2-C-13，N-15]亮氨酸。然后，我们通过C {N}旋转回波双共振（REDOR）测量C-13-N-15直接键。通过细胞壁双差直接测定具有连接到表面蛋白的桥的肽聚糖茎的增加（生物膜REDOR差减去浮游氏射流差异）。该程序消除了来自N-15同位素富集的差异的误差，以及从苏氨酸降解到甘氨酸的C-13标签的路由。对于浮游细胞和成熟的生物膜，20％的戊甘蔗桥不交联并且是潜在的表面蛋白质附着位点。这些位点中没有一个表面蛋白质，在浮游细胞中，但四分之一具有在成熟生物膜中附着的表面蛋白。此外，亮氨酸标记表明，富含亮氨酸的区域中的β股浓度在成熟的生物膜中加倍。因此，建立S.UUSTUS BIOFILM的主要事件是具有与细胞壁共价连接的表面蛋白质的细胞表面的大量装饰，并促进细胞 - 细胞粘附。

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