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首页> 美国卫生研究院文献>Infection and Immunity >Proteomic Analysis of Outer Membranes and Vesicles from Wild-Type Serogroup B Neisseria meningitidis and a Lipopolysaccharide-Deficient Mutant
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Proteomic Analysis of Outer Membranes and Vesicles from Wild-Type Serogroup B Neisseria meningitidis and a Lipopolysaccharide-Deficient Mutant

机译：野生型B群脑膜炎奈瑟氏球菌和脂多糖缺乏突变体的外膜和囊泡的蛋白质组学分析。

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摘要

Current experimental vaccines against serogroup B Neisseria meningitidis are based on meningococcal outer membrane (OM) proteins present in outer membrane vesicles (OMV) in which toxic lipopolysaccharide is depleted by detergent extraction. Knowledge of the composition of OM and OMV is essential for developing new meningococcal vaccines based on defined antigens. In the current study, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nanocapillary liquid chromatography-tandem mass spectrometry were used to investigate the proteomes of OM and OMV from meningococcal strain MC58 and OM from a lipopolysaccharide-deficient mutant. The analysis of OM revealed a composition that was much more complex than the composition that has been reported previously; a total of 236 proteins were identified, only 6.4% of which were predicted to be located in the outer membrane. The most abundant proteins included not only the well-established major OM proteins (PorA, PorB, Opc, Rmp, and Opa) but also other proteins, such as pilus-associated protein Q (PilQ) and a putative macrophage infectivity protein. All of these proteins were also present in OMV obtained by extraction of the OM with deoxycholate. There were markedly increased levels of some additional proteins in OM from the lipopolysaccharide-deficient mutant, including enzymes that contribute to the tricarboxylic acid cycle. In all the preparations, the proteins not predicted to have an OM location were predominantly periplasmic or cytoplasmic or had an unknown location, and relatively few cytoplasmic membrane proteins were detected. However, several proteins that have previously been identified as potential vaccine candidates were not detected in either OM preparations or in OMV. These results have important implications for the development and use of vaccines based on outer membrane proteins.
机译：当前针对血清群B脑膜炎奈瑟氏球菌的实验疫苗基于存在于外膜囊泡（OMV）中的脑膜炎球菌外膜（OM）蛋白，其中通过去污剂提取消耗了有毒的脂多糖。了解OM和OMV的组成对于开发基于确定抗原的新型脑膜炎球菌疫苗至关重要。在当前的研究中，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和纳米毛细管液相色谱-串联质谱法用于研究脑膜炎球菌MC58的OM和OMV蛋白质组以及脂多糖缺乏的突变体的OM。 OM的分析显示，其组成比以前报告的复杂得多。总共鉴定出236种蛋白质，其中只有6.4％被预测位于外膜中。最丰富的蛋白质不仅包括成熟的主要OM蛋白（PorA，PorB，Opc，Rmp和Opa），还包括其他蛋白质，如菌毛相关蛋白Q（PilQ）和假定的巨噬细胞感染性蛋白。所有这些蛋白质也存在于通过用脱氧胆酸盐提取OM获得的OMV中。脂多糖缺陷型突变体的OM中一些其他蛋白质的水平显着提高，包括有助于三羧酸循环的酶。在所有制剂中，未预测具有OM位置的蛋白质主要是周质或细胞质，或位置未知，并且检测到的胞质膜蛋白相对较少。但是，在OM制剂或OMV中均未检测到几种先前被鉴定为潜在疫苗候选物的蛋白质。这些结果对基于外膜蛋白的疫苗的开发和使用具有重要意义。

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