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Outer Membrane Vesicle Proteome of Porphyromonas gingivalis Is Differentially Modulated Relative to the Outer Membrane in Response to Heme Availability
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-05-29 , DOI: 10.1021/acs.jproteome.8b00153 Paul D. Veith 1 , Caroline Luong 1 , Kheng H. Tan 1 , Stuart G. Dashper 1 , Eric C. Reynolds 1
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-05-29 , DOI: 10.1021/acs.jproteome.8b00153 Paul D. Veith 1 , Caroline Luong 1 , Kheng H. Tan 1 , Stuart G. Dashper 1 , Eric C. Reynolds 1
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Porphyromonas gingivalis is an anaerobic, Gram-negative oral pathogen associated with chronic periodontitis. P. gingivalis has an obligate requirement for heme, which it obtains from the host. Heme availability has been linked to disease initiation and progression. In this study we used continuous culture of the bacterium to determine the effect of heme limitation and excess on the P. gingivalis proteome. Four biological replicates of whole cell lysate (WCL) and outer membrane vesicle (OMV) samples were digested with trypsin and analyzed by tandem mass spectrometry and MaxQuant label-free quantification. In total, 1211 proteins were quantified, with 108 and 49 proteins significantly changing in abundance more than 1.5-fold (p < 0.05) in the WCLs and OMVs, respectively. The proteins most upregulated in response to heme limitation were those involved in binding and transporting heme, whereas the four proteins most upregulated under the heme-excess condition constitute a putative heme efflux system. In general, the protein abundance ratios obtained for OMVs and WCLs agreed, indicating that changes to the OM protein composition are passed onto OMVs; however, 16 proteins were preferentially packaged into OMVs under one condition more than the other. In particular, moonlighting cytoplasmic proteins were preferentially associated with OMVs under heme excess.
中文翻译：
牙龈卟啉单胞菌的外膜囊泡蛋白质组相对于外膜对血红素可用性的差异调节。
牙龈卟啉单胞菌是与慢性牙周炎有关的厌氧，革兰氏阴性口腔病原体。牙龈卟啉单胞菌对血红素有强制性要求，它是从宿主那里获得的。血红素的可获得性与疾病的发生和发展有关。在这项研究中，我们使用细菌的连续培养来确定血红素限制和过量对牙龈卟啉单胞菌蛋白质组的影响。用胰蛋白酶消化全细胞裂解液（WCL）和外膜囊泡（OMV）样品的四个生物学重复样品，并通过串联质谱和MaxQuant-free定量分析。总共对1211种蛋白质进行了定量分析，其中108和49种蛋白质的丰度变化显着超过1.5倍（p<0.05）分别在WCL和OMV中。响应血红素限制而最上调的蛋白质是那些参与结合和转运血红素的蛋白质，而在血红素过量的条件下最上调的四种蛋白质构成了推测的血红素外排系统。通常，获得的OMV和WCL的蛋白质丰度比是一致的，这表明OM蛋白质组成的变化已传递到OMV上。然而，在一种条件下，优先将16种蛋白质包装到OMV中。特别是，在血红素过量的情况下，月光照下的细胞质蛋白优先与OMV相关。
更新日期：2018-05-30
中文翻译：
牙龈卟啉单胞菌的外膜囊泡蛋白质组相对于外膜对血红素可用性的差异调节。
牙龈卟啉单胞菌是与慢性牙周炎有关的厌氧，革兰氏阴性口腔病原体。牙龈卟啉单胞菌对血红素有强制性要求，它是从宿主那里获得的。血红素的可获得性与疾病的发生和发展有关。在这项研究中，我们使用细菌的连续培养来确定血红素限制和过量对牙龈卟啉单胞菌蛋白质组的影响。用胰蛋白酶消化全细胞裂解液（WCL）和外膜囊泡（OMV）样品的四个生物学重复样品，并通过串联质谱和MaxQuant-free定量分析。总共对1211种蛋白质进行了定量分析，其中108和49种蛋白质的丰度变化显着超过1.5倍（p<0.05）分别在WCL和OMV中。响应血红素限制而最上调的蛋白质是那些参与结合和转运血红素的蛋白质，而在血红素过量的条件下最上调的四种蛋白质构成了推测的血红素外排系统。通常，获得的OMV和WCL的蛋白质丰度比是一致的，这表明OM蛋白质组成的变化已传递到OMV上。然而，在一种条件下，优先将16种蛋白质包装到OMV中。特别是，在血红素过量的情况下，月光照下的细胞质蛋白优先与OMV相关。
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