Outer membrane vesicles from group B Neisseria meningitidis Δgna33 mutant: Proteomic and immunological comparison with detergent‐derived outer membrane vesicles

Abstract

We compared the proteome of detergent-derived group B Neisseria meningitidis (MenB) outer membrane vesicles (DOMVs) with the proteome of outer membrane vesicles (m-OMVs) spontaneously released into culture supernatant by MenB delta gna33, a mutant in which the gene coding for a lytic transglycosylase homologous to the E. coli MltA was deleted. In total, 138 proteins were identified in DOMVs by 1- and 2-DE coupled with MS; 64% of these proteins belonged to the inner membrane and cytoplasmic compartments. By contrast, most of the 60 proteins of m-OMVs were classified by PSORT as outer membrane proteins. When tested for their capacity to elicit bactericidal antibodies, m-OMVs elicited a broad protective activity against a large panel of MenB strains. Therefore, the identification of mutations capable of conferring an OMV-releasing phenotype in bacteria may represent an attractive approach to study bacterial membrane composition and organization, and to design new efficacious vaccine formulations.

Proteomic Characterization of the Whole Secretome of Legionella pneumophila and Functional Analysis of Outer Membrane Vesicles

“…Only 31 of 181 identified proteins (17%) were predicted to be cytoplasmic by both programs. This proportion is low compared to previous studies of OMV proteomes (30,80) and might have been the result of some cell lysis or cell autolysis that occurred during bacterial growth (76). When the list of proteins was searched, many type II and type IV secretion substrates were discovered.…”

“…A proteomic analysis of type II secreted effector proteins of L. pneumophila was recently described (26). The only proteomic studies of OMVs are the Neisseria meningitidis studies (30,80), which formed the basis for the development of MeNZB, an OMV vaccine against serogroup B (61).…”

Limitations of the rabbit pyrogen test for assessing meningococcal OMV based vaccines

“…The OMVs included in vaccines for human use are produced by detergent extraction (Bjune et al, 1991;de Moraes et al, 1992;Oster et al, 2005;Sierra et al, 1991), which removes lipoproteins and some LPS. However, a consequence of the detergent extraction is the carryover of cytoplasmic and periplasmic components (Ferrari et al, 2006;Vipond et al, 2006). Other known NEPs include porins, peptidoglycan, muramylpeptides and bacterial DNA (Hasiwa et al, 2013), all of which could conceivably be contained within OMV preparations either as genuine membrane components or as contaminants from the production process.…”

Significances of OMV and Extracellular Vesicle Proteomics

“…Minami et al, the changes in proteomic profiles of DRM fractions during cold acclimation using 2D-gel electrophoresis and mass analysis [134]. RND multidrug efflux pump membrane fusion proteins have been identified in the P. aeruginosa using membrane proteomics approach [86,106,[135][136][137]. Proteomic analyses of A. baumannii explain that histidine metabolism also have major role in the biofilm formation [138].…”