Yonezawa Hideo, Osaki Takako, Fukutomi Toshiyuki, Hanawa Tomoko, Kurata Satoshi, Zaman Cynthia, Hojo Fuhito, Kamiya Shigeru
Department of Infectious Diseases, Kyorin University School of Medicine, Tokyo, Japan
Department of Infectious Diseases, Kyorin University School of Medicine, Tokyo, Japan.
J Bacteriol. 2017 Feb 28;199(6). doi: 10.1128/JB.00729-16. Print 2017 Mar 15.
is one of the most common causes of bacterial infection in humans, and it forms biofilms on human gastric mucosal epithelium as well as on abiotic surfaces. Bacterial biofilm is critical not only for environmental survival but also for successful infection. We previously demonstrated that strain TK1402, which was isolated from a Japanese patient with duodenal and gastric ulcers, has high biofilm-forming ability relative to other strains. In addition, we showed that outer membrane vesicles (OMV) play an important role in biofilm formation. The aim of this study was to analyze which protein(s) in the OMV contributes to biofilm formation in TK1402. We obtained a spontaneous mutant strain derived from TK1402 lacking biofilm-forming ability. The protein profiles of the OMV were compared between this mutant strain and the wild type, and it was found that AlpB, an outer membrane protein in the OMV of the mutant strain, was markedly decreased compared to that of the wild type. Restoration of TK1402 to the mutant strain fully recovered the ability to form biofilm. However, restoration with from other strains demonstrated incomplete recovery of biofilm-forming ability. We therefore inferred that the variable region of AlpB (amino acid positions 121 to 146) was involved in TK1402 biofilm formation. In addition, diversification of the AlpB sequence was shown to affect the ability to adhere to AGS cells. These results demonstrate a new insight into the molecular mechanisms of host colonization by Bacterial biofilm is critical not only for environmental survival but also for successful infection. The mechanism of adherence to host cells mediated by cell surface adhesins has been the focus of many studies, but little is known regarding factors involved in biofilm formation. Our study demonstrated that AlpB plays an important role in biofilm formation and that this property depends upon the specific sequence of This in turn was shown to be important in the ability to adhere to gastric cells. We anticipate that these results will provide new insight into the molecular mechanisms of colonization.
是人类细菌感染最常见的原因之一,它可在人胃黏膜上皮以及非生物表面形成生物膜。细菌生物膜不仅对在环境中生存至关重要,对成功感染也很关键。我们之前证明,从一名患有十二指肠和胃溃疡的日本患者身上分离出的TK1402菌株,相对于其他菌株具有较高的生物膜形成能力。此外,我们还表明外膜囊泡(OMV)在生物膜形成中起重要作用。本研究的目的是分析OMV中的哪些蛋白质有助于TK1402中的生物膜形成。我们获得了一株源自TK1402的缺乏生物膜形成能力的自发突变菌株。比较了该突变菌株与野生型菌株OMV的蛋白质谱,发现突变菌株OMV中的一种外膜蛋白AlpB与野生型相比明显减少。将TK1402恢复到突变菌株可完全恢复生物膜形成能力。然而,用其他菌株的AlpB进行恢复显示生物膜形成能力未完全恢复。因此,我们推断AlpB的可变区(氨基酸位置121至146)参与了TK1402生物膜的形成。此外,AlpB序列的多样化被证明会影响对AGS细胞的粘附能力。这些结果揭示了对细菌生物膜宿主定植分子机制的新见解。细菌生物膜不仅对在环境中生存至关重要,对成功感染也很关键。由细胞表面粘附素介导的与宿主细胞粘附的机制一直是许多研究的重点,但关于生物膜形成所涉及的因素知之甚少。我们的研究表明AlpB在生物膜形成中起重要作用,并且这种特性取决于AlpB的特定序列。这反过来又被证明对粘附胃细胞的能力很重要。我们预计这些结果将为定植的分子机制提供新的见解。