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Preparation of Outer Membrane Vesicles from Myxococcus xanthus

James E. Berleman MZ Marcin Zemla JR Jonathan P Remis MA Manfred Auer*

0 Q&A 10645 Views Jan 20, 2016

Outer membrane vesicles (OMVs) represent a unique sub-cellular compartment of bacteria that may act as a scaffold for various extracellular activities, including intercellular signaling. Myxococcus xanthus (M. xanthus) is a predatory bacterium that engages in cell-cell behaviors such as fruiting body formation and contact dependent lysis of other microbes. The OMVs of M. xanthus have been shown to have an elaborate architecture of chains and tubes that can connect cells within a biofilm. These higher order OMV structures have been shown to contain proteins exchanged for community behaviors and small molecules that have antibiotic activities, and may help facilitate directed exchange. M. xanthus OMVs allow material transfer between neighboring cells for motility and predation.

Separation of Intracellular Vesicles for Immunoassays

TK Toshihiko Kobayashi* TT Tsubasa Tanaka NT Noriko Toyama-Sorimachi*

0 Q&A 11372 Views Aug 20, 2015

The endosome/lysosome systems play important roles in immune cell functions as signaling platforms. Immune cells utilize these endosome/lysosome for signal transduction or intercellular communication to elicit the proper immune responses, regulating the localization or the association of the signaling complexes. Here we introduce the procedures to separate the intracellular vesicles such as endosomes or lysosomes, which could be useful to identify the subcellular localization of the signaling complexes.

Vesicle Isolation from Bacillus subtilis Biofilm

Lisa Brown* AK Anne Kessler AC Arturo Casadevall

2 Q&A 12490 Views Mar 5, 2015

Bacterial biofilms are associated clinically with many bacterial infections including those caused by bacteria such as Pseudomonas aeruginosa and Staphylococcus aureus. In recent years, extracellular vesicles produced by bacteria have been isolated from biofilm communities. Vesicles have been described in depth and can encapsulate various virulence factors including toxins and immunomodulatory compounds. Vesicles may be important for virulence and survival by serving as a vehicle for the secretion and concentrated delivery of these molecules. Studying extracellular vesicles is an important step towards understanding biofilm formation, structure, and disruption with the ultimate goal of preventing or treating hospital infections caused by bacterial pathogens residing in biofilms. Here we describe the protocol for isolating vesicles from biofilm produced by Bacillus subtilis.

Preparation of Outer Membrane Vesicle from Escherichia coli

Oh Youn Kim BH Bok Sil Hong KP Kyong-Su Park YY Yae Jin Yoon SC Seng Jin Choi WL Won Hee Lee TR Tae-Young Roh YK Yoon-Keun Kim

Yong Song Gho*

3 Q&A 21581 Views Dec 5, 2013

Outer membrane vesicles (OMVs) are spherical bilayered phospholipids of 20-200 nm in size produced from all Gram-negative bacteria and Gram-positive bacteria investigated to date. Previous biochemical and proteomic studies have revealed that the Gram-negative bacteria-derived OMVs are composed of various components like outer membrane proteins, lipopolysaccharides, outer membrane lipids, periplasmic proteins, DNA, and RNA. Here, in this protocol, we describe the method to isolate the OMVs from the culture supernatant of Escherichia coli (E. coli).

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