Flagella proteins contribute to the production of outer membrane vesicles from Escherichia coli W3110
Flagella proteins contribute to the production of outer membrane vesicles from Escherichia coli W3110
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DOI：
10.1016/j.bbrc.2013.10.022
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发表时间：
2013-11-08
影响因子：
3.1
通讯作者：
Kawasaki, Kiyoshi
中科院分区：
文献类型：
--
作者：
Manabe, Takayuki;Kato, Mayu;Kawasaki, Kiyoshi
包括大肠杆菌在内的革兰氏阴性细菌会释放源自细菌外膜的外膜囊泡 (OMV)。 OMV 通过将成分递送到细菌细胞外的位置来促进细菌细胞间的通讯和宿主微生物的相互作用。为了探索 OMV 生物合成中涉及的分子机制，研究了一种主要 OMV 蛋白在大肠杆菌 W3110（广泛使用的标准大肠杆菌 K-12 菌株）生产 OMV 过程中的作用。除了用作 OMV 标记蛋白的 OmpC 和 OmpA 之外，对大肠杆菌 W3110 OMV 的分析表明，它们还含有丰富水平的 FliC（也称为鞭毛蛋白）。膜不可渗透的生物素标记试剂不会在完整的 OMV 中标记 FliC，但在声波破坏的 OMV 中标记 FliC，表明 FliC 位于 OMV 的管腔中。与表达野生型 fliC 的亲本菌株相比，具有 fliC 无效突变的大肠杆菌菌株产生的 OMV 量在蛋白质和磷酸盐水平上均有所减少。此外，来自大肠杆菌W3110的菌株flgK（编码鞭毛钩相关蛋白，与FliC一起对鞭毛合成至关重要）具有无效突变，也比亲本菌株产生更少的OMV。综上所述，这些结果表明形成鞭毛的能力，包括鞭毛蛋白的合成，影响大肠杆菌 W3110 OMV 的产生。 (C) 2013 Elsevier Inc. 保留所有权利。
Gram-negative bacteria, including Escherichia coli, release outer membrane vesicles (OMVs) that are derived from the bacterial outer membrane. OMVs contribute to bacterial cell-cell communications and host-microbe interactions by delivering components to locations outside the bacterial cell. In order to explore the molecular machinery involved in OMV biogenesis, the role of a major OMV protein was examined in the production of OMVs from E. coli W3110, which is a widely used standard E. coli K-12 strain. In addition to OmpC and OmpA, which are used as marker proteins for OMVs, an analysis of E. coli W3110 OMVs revealed that they also contain abundant levels of FliC, which is also known as flagellin. A membrane-impermeable biotin-labeling reagent did not label FliC in intact OMVs, but labeled FliC in sonically disrupted OMVs, suggesting that FliC is localized in the lumen of OMV. Compared to the parental strain expressing wild-type fliC, an E. coli strain with a fliC-null mutation produced reduced amounts of OMVs based on both protein and phosphate levels. In addition, an E. coli W3110-derived strain with a null-mutation in flgK, which encodes flagellar hook-associated protein that is essential along with FliC for flagella synthesis, also produced fewer OMVs than the parental strain. Taken together, these results indicate that the ability to form flagella, including the synthesis of flagella proteins, affects the production of E. coli W3110 OMVs. (C) 2013 Elsevier Inc. All rights reserved.