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经 2D 电泳和质谱分析,中间普氏菌在氧化应激后的差异调节蛋白。

Differentially regulated proteins in Prevotella intermedia after oxidative stress analyzed by 2D electrophoresis and mass spectrometry.

机构信息

Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos, 6627, Pampulha, CEP 31.270-901, Belo Horizonte, MG, Brazil.

出版信息

Anaerobe. 2012 Feb;18(1):76-82. doi: 10.1016/j.anaerobe.2011.12.008. Epub 2011 Dec 16.

Abstract

Prevotella intermedia is a rod-shaped, Gram-negative anaerobic bacterium found in human indigenous microbiota that plays an important role in opportunistic infections. The successful colonization depends on the ability of anaerobes to respond to oxidative stress (OS) in oxygenated tissues as well as to resist oxidative events from the host immune system until anaerobic conditions are present at the infection site. As knowledge of the mechanisms of protection against OS in Prevotella is limited, studies are needed to clarify aspects of molecular biology, physiology and ecology of this bacterium. The aim of this study was to access the proteins differentially regulated in P. intermedia after exposure to molecular oxygen by using two-dimensional gel electrophoresis (2DE) associated with the approach of MALDI-TOF/TOF Tandem Mass Spectrometry. The identity of the protein was evaluated by database search for homologous genomic sequences of P. intermedia strain 17 (TIGR). Twenty five out of 72 proteins found were identified as up-regulated (17) or down-regulated (9). These proteins were related to a variety of metabolic process, some of which could be associated to antioxidant and redox regulatory roles. Our data indicate that OS may stimulate an adaptive response in P. intermedia whose effect on its biology may be evidenced by the increase in aerotolerance and changes in protein abundance in the oxygen adapted cells.

摘要

中间普雷沃氏菌是一种杆状、革兰氏阴性的厌氧菌,存在于人体固有微生物群中,在机会性感染中发挥重要作用。成功定植取决于厌氧菌在含氧组织中对氧化应激(OS)的反应能力,以及在感染部位出现厌氧条件之前抵抗来自宿主免疫系统的氧化事件的能力。由于对中间普雷沃氏菌抵御 OS 的机制知之甚少,因此需要研究来阐明该细菌的分子生物学、生理学和生态学的各个方面。本研究旨在通过二维凝胶电泳(2DE)结合 MALDI-TOF/TOF 串联质谱技术,研究中间普雷沃氏菌在暴露于分子氧后的差异调节蛋白。通过对中间普雷沃氏菌 17 株(TIGR)的同源基因组序列进行数据库搜索,评估了该蛋白的身份。在 72 种发现的蛋白中,有 25 种被鉴定为上调(17 种)或下调(9 种)。这些蛋白与多种代谢过程有关,其中一些可能与抗氧化和氧化还原调节作用有关。我们的数据表明,OS 可能会刺激中间普雷沃氏菌产生适应性反应,其对生物学的影响可能表现在耐氧性增加和适应氧气的细胞中蛋白丰度的变化。

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