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Outer membrane vesicles (OMVs) are vesicles released from the outer membranes of Gram-negative bacteria. While Gram-positive bacteria release vesicles as well, those vesicles fall under the broader category of bacterial membrane vesicles (MVs). OMVs were the first MVs to be discovered, and are distinguished from outer inner membrane vesicles (OIMVs), which are gram-negative bacterial vesicles containing portions of both the outer and inner bacterial membrane. [1] Outer membrane vesicles were first discovered and characterized using transmission-electron microscopy [2] by Indian Scientist Prof. Smriti Narayan Chatterjee and J. Das in 1966-67. [3] [4] OMVs are ascribed the functionality to provide a manner to communicate among themselves, with other microorganisms in their environment and with the host. These vesicles are involved in trafficking bacterial cell signaling biochemicals, which may include DNA, RNA, proteins, endotoxins and allied virulence molecules. This communication happens in microbial cultures in oceans, [5] inside animals, plants and even inside the human body. [6]
Gram-negative bacteria deploy their periplasm to secrete OMVs for trafficking bacterial biochemicals to target cells in their environment. OMVs also can carry endotoxic lipopolysaccharide that may contribute to disease processes in their host. [7] [8] This mechanism imparts a variety of benefits like, long-distance delivery of bacterial secretory cargo with minimized hydrolytic degradation and extra-cellular dilution, also supplemented with other supportive molecules (e.g., virulence factors) to accomplish a specific job and yet, keeping a safe-distance from the defense arsenal of the targeted cells. Biochemical signals trafficked by OMVs may vary largely during 'war and peace' situations. In 'complacent' bacterial colonies, OMVs may be used to carry DNA to 'related' microbes for genetic transformations, and also translocate cell signaling molecules for quorum sensing and biofilm formation. During 'challenge' from other cell types around, OMVs may be preferred to carry degradation and subversion enzymes. Likewise, OMVs may contain more of invasion proteins at the host–pathogen interface (Fig. 1). It is expected that environmental factors around the secretory microbes are responsible for inducing these bacteria to synthesize and secrete specifically-enriched OMVs, physiologically suiting the immediate task. Thus, bacterial OMVs, being strong immunomodulators, [9] can be manipulated for their immunogenic contents and utilized as potent pathogen-free vaccines [10] for immunizing humans and animals against threatening infections. VA-MENGOC-BC and Bexsero against meningitis are currently the only OMV vaccines approved in the US, though an OMV vaccine for gonorrhea is seeking approval. [11] [12]
Gram-negative bacteria have a double set of lipid bilayers. An inner bilayer, the inner cell membrane, encloses the cytoplasm or cytosol. Surrounding this inner cell membrane there is a second bilayer called the bacterial outer membrane. The compartment or space between these two membranes is called the periplasm or periplasmic space. In addition, there is a firm cell wall consisting of peptidoglycan layer, which surrounds the cell membrane and occupies the periplasmic space. The peptidoglycan layer provides some rigidity for maintaining the bacterial cell shape, besides also protecting the microbe against challenging environments.
The first step in biogenesis of gram-negative bacterial OMVs, [13] is bulging of outer membrane above the peptidoglycan layer. Accumulation of phospholipids in the outside of the outer membrane is thought to be the basis of this outwards bulging of the outer membrane. [14] This accumulation of phospholipids can be regulated by the VacJ/Yrb ABC transport system that transfers phospholipids from the outside of OM to the inner side. [14] Additionally, environmental conditions as sulfur depletion can trigger a state of phospholipid overproduction that causes increased OMV release. [15]
The actual release of the vesicle from the outer membrane remains unclear. It is likely that vesicle structures can be released spontaneously. Alternatively, it has been suggested that few proteins 'rivet' the outer and cell membranes together, so that the periplasmic bulge protrudes like a 'ballooned' pocket of inflated periplasm out from the surface of the outer membrane. Lateral diffusion of 'rivet complexes' may help in pinching off large bulges of periplasm as OMVs. [16]
Bacterial membrane vesicles' dispersion along the cell surface was measured in live Escherichia coli , commensal bacteria common in the human gut. Antibiotic treatment altered vesicle dynamics, vesicle-to-membrane affinity, and surface properties of the cell membranes, generally enhancing vesicle transport along the surfaces of bacterial membranes and suggesting that their motion properties could be a signature of antibiotic stress. [17] Despite this first high-resolution, quantitative tracking of bacterial OMVs, detailed experimental work is still awaited to understand the biomechanics of OMV biogenesis and transport. OMVs are also under focus of current research in exocytosis in prokaryotes via outer membrane vesicle trafficking for intra-species, inter-species and inter-kingdom cell signaling, which is slated to change our mindset on virulence of microbes, host–pathogen interactions and inter-relationships among variety of species in earth's ecosystem.
Gram stain, is a method of staining used to classify bacterial species into two large groups: gram-positive bacteria and gram-negative bacteria. It may also be used to diagnose a fungal infection. The name comes from the Danish bacteriologist Hans Christian Gram, who developed the technique in 1884.
In bacteriology, gram-positive bacteria are bacteria that give a positive result in the Gram stain test, which is traditionally used to quickly classify bacteria into two broad categories according to their type of cell wall.
Gram-negative bacteria are bacteria that unlike gram-positive bacteria do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. Their defining characteristic is their cell envelope, which consists of a thin peptidoglycan cell wall sandwiched between an inner (cytoplasmic) membrane and an outer membrane. These bacteria are found in all environments that support life on Earth.
Peptidoglycan or murein is a unique large macromolecule, a polysaccharide, consisting of sugars and amino acids that forms a mesh-like layer (sacculus) that surrounds the bacterial cytoplasmic membrane. The sugar component consists of alternating residues of β-(1,4) linked N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM). Attached to the N-acetylmuramic acid is an oligopeptide chain made of three to five amino acids. The peptide chain can be cross-linked to the peptide chain of another strand forming the 3D mesh-like layer. Peptidoglycan serves a structural role in the bacterial cell wall, giving structural strength, as well as counteracting the osmotic pressure of the cytoplasm. This repetitive linking results in a dense peptidoglycan layer which is critical for maintaining cell form and withstanding high osmotic pressures, and it is regularly replaced by peptidoglycan production. Peptidoglycan hydrolysis and synthesis are two processes that must occur in order for cells to grow and multiply, a technique carried out in three stages: clipping of current material, insertion of new material, and re-crosslinking of existing material to new material.
Bdellovibrio is a genus of Gram-negative, obligate aerobic bacteria. One of the more notable characteristics of this genus is that members can prey upon other Gram-negative bacteria and feed on the biopolymers, e.g. proteins and nucleic acids, of their hosts. They have two lifestyles: a host-dependent, highly mobile phase, the "attack phase", in which they form "bdelloplasts" in their host bacteria; and a slow-growing, irregularly shaped, host-independent form.
Treponema pallidum, formerly known as Spirochaeta pallida, is a microaerophilic spirochaete bacterium with subspecies that cause the diseases syphilis, bejel, and yaws. It is known to be transmitted only among humans and baboons. It is a helically coiled microorganism usually 6–15 μm long and 0.1–0.2 μm wide. T. pallidum's lack of both a tricarboxylic acid cycle and processes for oxidative phosphorylation results in minimal metabolic activity. The treponemes have cytoplasmic and outer membranes. Using light microscopy, treponemes are visible only by using dark-field illumination. T. pallidum consists of three subspecies, T. p. pallidum, T. p. endemicum, and T. p. pertenue, each of which has a distinct associated disease.
Secretion is the movement of material from one point to another, such as a secreted chemical substance from a cell or gland. In contrast, excretion is the removal of certain substances or waste products from a cell or organism. The classical mechanism of cell secretion is via secretory portals at the plasma membrane called porosomes. Porosomes are permanent cup-shaped lipoprotein structures embedded in the cell membrane, where secretory vesicles transiently dock and fuse to release intra-vesicular contents from the cell.
The periplasm is a concentrated gel-like matrix in the space between the inner cytoplasmic membrane and the bacterial outer membrane called the periplasmic space in gram-negative bacteria. Using cryo-electron microscopy it has been found that a much smaller periplasmic space is also present in gram-positive bacteria, between cell wall and the plasma membrane. The periplasm may constitute up to 40% of the total cell volume of gram-negative bacteria, but is a much smaller percentage in gram-positive bacteria.
The cell envelope comprises the inner cell membrane and the cell wall of a bacterium. In Gram-negative bacteria an outer membrane is also included. This envelope is not present in the Mollicutes where the cell wall is absent.
Braun's lipoprotein, found in some gram-negative cell walls, is one of the most abundant membrane proteins; its molecular weight is about 7.2 kDa. It is bound at its C-terminal end by a covalent bond to the peptidoglycan layer and is embedded in the outer membrane by its hydrophobic head. BLP tightly links the two layers and provides structural integrity to the outer membrane.
Moraxella catarrhalis is a fastidious, nonmotile, Gram-negative, aerobic, oxidase-positive diplococcus that can cause infections of the respiratory system, middle ear, eye, central nervous system, and joints of humans. It causes the infection of the host cell by sticking to the host cell using trimeric autotransporter adhesins.
The bacterial outer membrane is found in gram-negative bacteria. Gram-negative bacteria form two lipid bilayers in their cell envelopes - an inner membrane (IM) that encapsulates the cytoplasm, and an outer membrane (OM) that encapsulates the periplasm.
A bacterium, despite its simplicity, contains a well-developed cell structure which is responsible for some of its unique biological structures and pathogenicity. Many structural features are unique to bacteria and are not found among archaea or eukaryotes. Because of the simplicity of bacteria relative to larger organisms and the ease with which they can be manipulated experimentally, the cell structure of bacteria has been well studied, revealing many biochemical principles that have been subsequently applied to other organisms.
In molecular biology, an autotransporter domain is a structural domain found in some bacterial outer membrane proteins. The domain is always located at the C-terminal end of the protein and forms a beta-barrel structure. The barrel is oriented in the membrane such that the N-terminal portion of the protein, termed the passenger domain, is presented on the cell surface. These proteins are typically virulence factors, associated with infection or virulence in pathogenic bacteria.
The cell membrane is a biological membrane that separates and protects the interior of a cell from the outside environment. The cell membrane consists of a lipid bilayer, made up of two layers of phospholipids with cholesterols interspersed between them, maintaining appropriate membrane fluidity at various temperatures. The membrane also contains membrane proteins, including integral proteins that span the membrane and serve as membrane transporters, and peripheral proteins that loosely attach to the outer (peripheral) side of the cell membrane, acting as enzymes to facilitate interaction with the cell's environment. Glycolipids embedded in the outer lipid layer serve a similar purpose. The cell membrane controls the movement of substances in and out of a cell, being selectively permeable to ions and organic molecules. In addition, cell membranes are involved in a variety of cellular processes such as cell adhesion, ion conductivity, and cell signalling and serve as the attachment surface for several extracellular structures, including the cell wall and the carbohydrate layer called the glycocalyx, as well as the intracellular network of protein fibers called the cytoskeleton. In the field of synthetic biology, cell membranes can be artificially reassembled.
Membrane vesicle trafficking in eukaryotic animal cells involves movement of biochemical signal molecules from synthesis-and-packaging locations in the Golgi body to specific release locations on the inside of the plasma membrane of the secretory cell. It takes place in the form of Golgi membrane-bound micro-sized vesicles, termed membrane vesicles (MVs).
The type 2 secretion system is a type of protein secretion machinery found in various species of Gram-negative bacteria, including many human pathogens such as Pseudomonas aeruginosa and Vibrio cholerae. The type II secretion system is one of six protein secretory systems commonly found in Gram-negative bacteria, along with the type I, type III, and type IV secretion systems, as well as the chaperone/usher pathway, the autotransporter pathway/type V secretion system, and the type VI secretion system. Like these other systems, the type II secretion system enables the transport of cytoplasmic proteins across the lipid bilayers that make up the cell membranes of Gram-negative bacteria. Secretion of proteins and effector molecules out of the cell plays a critical role in signaling other cells and in the invasion and parasitism of host cells.
BamA is a β-barrel, outer membrane protein found in Gram-negative bacteria and it is the main and vital component of the β-barrel assembly machinery (BAM) complex in those bacteria. BAM Complex consists of five components; BamB, BamC, BamD, BamE and BamA. This complex is responsible in catalyzing folding and insertion of β-barrel proteins into the outer membrane of Gram-negative bacteria.
Bacterial secretion systems are protein complexes present on the cell membranes of bacteria for secretion of substances. Specifically, they are the cellular devices used by pathogenic bacteria to secrete their virulence factors to invade the host cells. They can be classified into different types based on their specific structure, composition and activity. Generally, proteins can be secreted through two different processes. One process is a one-step mechanism in which proteins from the cytoplasm of bacteria are transported and delivered directly through the cell membrane into the host cell. Another involves a two-step activity in which the proteins are first transported out of the inner cell membrane, then deposited in the periplasm, and finally through the outer cell membrane into the host cell.
Darobactin is an experimental antibiotic compound that may be effective against Gram-negative bacteria. If it can be developed into a human-compatible form it would be the first to come from an animal microbiome.