Flagella proteins contribute to the production of outer membrane vesicles from Escherichia coli W3110 |
| |
Authors: | Takayuki Manabe Mayu KatoTakayuki Ueno Kiyoshi Kawasaki |
| |
Affiliation: | Faculty of Pharmaceutical Sciences, Doshisha Women’s College, Kyotanabe, Kyoto 610-0395, Japan |
| |
Abstract: | 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. |
| |
Keywords: | OMVs, outer membrane vesicles LPS, lipopolysaccharide SDS, sodium dodecyl sulfate PAGE, polyacrylamide gel electrophoresis MALDI-TOF, matrix-assisted laser desorption ionization&ndash time of flight |
本文献已被 ScienceDirect 等数据库收录! |
|