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Localization of Outer Membrane Proteins in Treponema denticola by Quantitative Proteome Analyses of Outer Membrane Vesicles and Cellular Fractions.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-02-26 , DOI: 10.1021/acs.jproteome.8b00860
Paul D Veith 1 , Michelle D Glew 1 , Dhana G Gorasia 1 , Dina Chen 1 , Neil M O'Brien-Simpson 1 , Eric C Reynolds 1
Affiliation  

The identification and localization of outer membrane proteins (Omps) and lipoproteins in pathogenic treponemes such as T. denticola (periodontitis) and T. pallidum (syphilis) has been challenging. In this study, label-free quantitative proteomics using MaxQuant was applied to naturally produced outer membrane vesicles (OMVs) and cellular fractions to identify 1448 T. denticola proteins. Of these, 90 proteins were localized to the outer membrane (OM) comprising 59 lipoproteins, 25 β-barrel proteins, and six other putative OM-associated proteins. Twenty-eight lipoproteins were localized to the inner membrane (IM), and 43 proteins were assigned to the periplasm. The signal cleavage regions of the OM and IM lipoprotein sequences were different and may reveal the signals for their differential localization. Proteins significantly enriched in OMVs included dentilisin, proteins containing leucine-rich repeats, and several lipoproteins containing FGE-sulfatase domains. Blue native PAGE analysis enabled the native size of the dentilisin complex and Msp to be determined and revealed that the abundant β-barrel Omps TDE2508 and TDE1717 formed large complexes. In addition to the large number of integral Omps and potentially surface-located lipoproteins identified in T. denticola, many such proteins were also newly identified in T. pallidum through homology, generating new targets for vaccine development in both species.

中文翻译:

通过外膜囊泡和细胞组分的定量蛋白质组分析,在密闭螺旋体中外膜蛋白的定位。

在致病性色氨酸如牙本质衣原体(牙周炎)和苍白衣原体(梅毒)中,外膜蛋白(Omps)和脂蛋白的鉴定和定位一直具有挑战性。在这项研究中,使用MaxQuant的无标签定量蛋白质组学被应用于天然产生的外膜囊泡(OMV)和细胞级分,以鉴定1448种​​树突触杆菌蛋白质。其中90种蛋白质位于外膜(OM)上,外膜(OM)包含59种脂蛋白,25种β-barrel蛋白和6种其他假定的OM相关蛋白。28个脂蛋白位于内膜(IM),43种蛋白被分配至周质。OM和IM脂蛋白序列的信号切割区域是不同的,并且可能揭示其差异定位的信号。富含OMV的蛋白质包括牙本质素,含有富亮氨酸重复序列的蛋白质和含有FGE硫酸酯酶结构域的几种脂蛋白。蓝色天然PAGE分析能够确定牙本质素复合物和Msp的天然大小,并揭示了丰富的β-桶形Omps TDE2508和TDE1717形成了大的复合物。除了在鼠李糖单胞菌中鉴定出大量完整的Omps和可能位于表面的脂蛋白外,还通过同源性在梅毒螺旋体中新鉴定了许多这样的蛋白质,从而为这两个物种的疫苗开发提供了新的靶标。蓝色天然PAGE分析能够确定牙本质素复合物和Msp的天然大小,并揭示了丰富的β-桶形Omps TDE2508和TDE1717形成了大的复合物。除了在鼠李糖单胞菌中鉴定出大量完整的Omps和可能位于表面的脂蛋白外,还通过同源性在梅毒螺旋体中新鉴定了许多这样的蛋白质,从而为这两个物种的疫苗开发提供了新的靶标。蓝色天然PAGE分析能够确定牙本质素复合物和Msp的天然大小,并揭示了丰富的β-桶形Omps TDE2508和TDE1717形成了大的复合物。除了在鼠李糖单胞菌中鉴定出大量完整的Omps和可能位于表面的脂蛋白外,还通过同源性在梅毒螺旋体中新鉴定了许多这样的蛋白质,从而为这两个物种的疫苗开发提供了新的靶标。
更新日期:2019-02-27
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