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基于蛋白质组学对牙龈卟啉单胞菌中抗氧化应激系统的分析

Proteomics-based analysis of a counter-oxidative stress system in Porphyromonas gingivalis.

作者信息

Okano Soichiro, Shibata Yasuko, Shiroza Teruaki, Abiko Yoshimitsu

机构信息

Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, 2-870-1 Sakaecho-nishi, Matsudo, Chiba 271-8587 Chiba, Japan.

出版信息

Proteomics. 2006 Jan;6(1):251-8. doi: 10.1002/pmic.200401338.

Abstract

Porphyromonas gingivalis is a Gram-negative anaerobic pathogen associated with chronic periodontitis. Although anaerobic, P. gingivalis exhibits a high degree of aerotolerance, which enables it to survive within periodontal pockets. The aim of the present study was to examine the effect of oxidative stress on protein expression in P. gingivalis to obtain a better understanding of the mechanism underlying its aerotolerance. To accomplish this, P. gingivalis cells were grown under conditions of hemin limitation (0.01 microg/mL) to avoid the oxygen protective effect of hemin on oxidative stress. The proteins were then extracted from cultures either left untreated or subjected to oxidative stress and separated by 2-DE. The resultant protein expression profiles were examined by image scanning, and those found to differ depending on the presence or absence of aeration were subjected to MALDI-MS and then analyzed using the ORF database of P. gingivalis W83 from The Institute of Genomic Research. Oxidative stress was found to affect the expression of numerous proteins in P. gingivalis cells. In particular, the levels of HtpG, GroEL, DnaK, AhpC, TPR domain protein, and trigger factor were substantially increased.

摘要

牙龈卟啉单胞菌是一种与慢性牙周炎相关的革兰氏阴性厌氧病原体。尽管牙龈卟啉单胞菌是厌氧菌,但它表现出高度的耐氧性,这使其能够在牙周袋内存活。本研究的目的是研究氧化应激对牙龈卟啉单胞菌蛋白质表达的影响,以便更好地了解其耐氧性的潜在机制。为实现这一目的,牙龈卟啉单胞菌细胞在血红素限制条件(0.01微克/毫升)下培养,以避免血红素对氧化应激的氧保护作用。然后从未经处理或遭受氧化应激的培养物中提取蛋白质,并通过双向电泳进行分离。通过图像扫描检查所得的蛋白质表达谱,将那些根据是否通气而发现有差异的蛋白质进行基质辅助激光解吸电离质谱分析,然后使用来自基因组研究所的牙龈卟啉单胞菌W83的开放阅读框数据库进行分析。发现氧化应激会影响牙龈卟啉单胞菌细胞中许多蛋白质的表达。特别是,热激蛋白90、热激蛋白60、热激蛋白70、烷基过氧化氢还原酶C、四肽重复结构域蛋白和触发因子的水平大幅增加。

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