A Cellular Deficiency of Gangliosides Causes Hypersensitivity to Clostridium perfringens Phospholipase C

Flores-Dı́az, Marietta; Alape-Girón, Alberto; Clark, Graeme C.; Catimel, Bruno; Hirabayashi, Yoshio; Nice, Edouard C.; Gutiérrez, José M.; Titball, Richard W.; Thelestam, Mónica

doi:10.1074/jbc.m500278200

Cited by 58 publications

(53 citation statements)

References 63 publications

(91 reference statements)

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“…Alpha-toxin is an essential virulence factor in gas gangrene (2), and perfringolysin O, although not essential, has been found to have a synergistic role with alpha-toxin, enhancing the disease process (3). Synergy between alpha-toxin and the NanI sialidase was also observed in experiments that showed that purified alpha-toxin had greater pathological effects on cultured cell lines that had been pretreated with NanI (8). Inoculation of mice with both purified alpha-toxin and NanI resulted in increased levels of plasma creatine kinase, a marker for muscle necrosis, compared to the levels after inoculation of alpha-toxin alone (8).…”

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confidence: 98%

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The NanI and NanJ Sialidases of Clostridium perfringens Are Not Essential for Virulence

et al. 2009

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The essential toxin in Clostridium perfringens-mediated gas gangrene or clostridial myonecrosis is alphatoxin, although other toxins and extracellular enzymes may also be involved. In many bacterial pathogens extracellular sialidases are important virulence factors, and it has been suggested that sialidases may play a role in gas gangrene. C. perfringens strains have combinations of three different sialidase genes, two of which, nanI and nanJ, encode secreted sialidases. The nanI and nanJ genes were insertionally inactivated by homologous recombination in derivatives of sequenced strain 13 and were shown to encode two functional secreted sialidases, NanI and NanJ. Analysis of these derivatives showed that NanI was the major sialidase in this organism. Mutation of nanI resulted in loss of most of the secreted sialidase activity, and the residual activity was eliminated by subsequent mutation of the nanJ gene. Only a slight reduction in the total sialidase activity was observed in a nanJ mutant. Cytotoxicity assays using the B16 melanoma cell line showed that supernatants containing NanI or overexpressing NanJ enhanced alpha-toxin-mediated cytotoxicity. Finally, the ability of nanI, nanJ, and nanIJ mutants to cause disease was assessed in a mouse myonecrosis model. No attenuation of virulence was observed for any of these strains, providing evidence that neither the NanI sialidase nor the NanJ sialidase is essential for virulence.Clostridium perfringens type A is the causative agent of human gas gangrene, or clostridial myonecrosis, and human food poisoning (25,27). It produces many secreted hydrolytic enzymes and toxins, including alpha-toxin and perfringolysin O. C. perfringens strains can also encode up to three sialidases, but the three sialidase genes, nanH, nanI, and nanJ, are not present in all of the strains that have been completely sequenced. Strain ATCC 13124 encodes all three sialidases (18), while strain 13 encodes both of the large sialidases, NanI and NanJ, but not the smaller NanH enzyme (32). The food poisoning isolate SM101 encodes NanH but not NanI or NanJ (18).Sialidases have been implicated in the virulence of several bacterial pathogens. They have been shown to enhance the pathogenesis of disease through synergistic effects with other bacterial factors. For example, Vibrio cholerae sialidase enhances the activity of cholera toxin (10), Pseudomonas aeruginosa sialidase increases the binding of this organism to the cells of susceptible patients (6), and the two sialidases of Streptococcus pneumoniae contribute to the progression of infection in several animal models (16,23,37). More recently, a surfaceexposed sialidase was shown to be required for persistence of the canine pathogen Capnophagia canimorsus (15). Alpha-toxin is an essential virulence factor in gas gangrene (2), and perfringolysin O, although not essential, has been found to have a synergistic role with alpha-toxin, enhancing the disease process (3). Synergy between alpha-toxin and the NanI sialidase was also observed in experiment...

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confidence: 98%

“…As a result of its location, NanI may also interact with the extracellular environment of the host tissue during infection. In addition to its synergy with alpha-toxin (8), NanI has also been shown to have synergistic effects with ε-toxin (31), which is required for C. perfringens type B-and D-mediated diseases (34,39,40).…”

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The NanI and NanJ Sialidases of Clostridium perfringens Are Not Essential for Virulence

et al. 2009

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“…C. perfringens alpha-toxin-induced myotoxicity and lethality are at least partially mediated by ROS, as myonecrosis and mortality in a murine model of gas gangrene are reduced by free radical scavengers (222). C. perfringens alpha-toxin induces cell death in various cell types, particularly those with low ganglioside content in their plasma membrane (223). Muscle has the lowest concentration of complex gangliosides among mammalian tissues, offering a possible explanation for the high susceptibility of muscle fibers to alpha-toxin (223).…”

Section: Phospholipase Csmentioning

confidence: 99%

“…C. perfringens alpha-toxin induces cell death in various cell types, particularly those with low ganglioside content in their plasma membrane (223). Muscle has the lowest concentration of complex gangliosides among mammalian tissues, offering a possible explanation for the high susceptibility of muscle fibers to alpha-toxin (223). Other clostridial PLCs with a structure similar to C. perfringens alpha-toxin display lower toxicities due to different amino acid substitutions (212,224).…”

Section: Phospholipase Csmentioning

confidence: 99%

Bacterial Sphingomyelinases and Phospholipases as Virulence Factors

Flores-Dı́az

Monturiol-Gross

Naylor

et al. 2016

Microbiol Mol Biol Rev

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SUMMARYBacterial sphingomyelinases and phospholipases are a heterogeneous group of esterases which are usually surface associated or secreted by a wide variety of Gram-positive and Gram-negative bacteria. These enzymes hydrolyze sphingomyelin and glycerophospholipids, respectively, generating products identical to the ones produced by eukaryotic enzymes which play crucial roles in distinct physiological processes, including membrane dynamics, cellular signaling, migration, growth, and death. Several bacterial sphingomyelinases and phospholipases are essential for virulence of extracellular, facultative, or obligate intracellular pathogens, as these enzymes contribute to phagosomal escape or phagosomal maturation avoidance, favoring tissue colonization, infection establishment and progression, or immune response evasion. This work presents a classification proposal for bacterial sphingomyelinases and phospholipases that considers not only their enzymatic activities but also their structural aspects. An overview of the main physiopathological activities is provided for each enzyme type, as are examples in which inactivation of a sphingomyelinase- or a phospholipase-encoding gene impairs the virulence of a pathogen. The identification of sphingomyelinases and phospholipases important for bacterial pathogenesis and the development of inhibitors for these enzymes could generate candidate vaccines and therapeutic agents, which will diminish the impacts of the associated human and animal diseases.

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“…Control vesicles, i.e., those containing PC:SM:PE:Chol (in equimolar proportions) become stained at a somewhat higher rate than those containing DAG and/or Cer in addition. Note that phase characterization in an identical system ( 22 ) and by the various important early ( 30,32,57 ) and contemporary ( 42,49,(58)(59)(60) studies of lipases and lipid domains that have been carried out using natural lipids. GUV are examined individually in most cases, without contact with other vesicles and under reduced mobility conditions; thus, vesicle aggregation or fusion cannot be observed under those conditions.…”

Section: Enzyme Activity On Luvmentioning

confidence: 99%

Imaging the early stages of phospholipase C/sphingomyelinase activity on vesicles containing coexisting ordered-disordered and gel-fluid domains

Ibarguren

López

Montes

et al. 2011

Journal of Lipid Research

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Supplementary key words ceramides • cholesterol • diacylglycerol • fl uorescence microscopy • lipid rafts • membranes/physical chemistry • sphingolipidsPhospholipases are essential enzymes in maintaining membrane homeostasis and in the generation of metabolic signals. They are also powerful tools that bacteria use to infect and eventually destroy eukaryotic cells by helping in the degradation of the target cell membranes. Phospholipase C (PLC) enzymes cleave the phosphodiester bond between the diacylglycerol moiety and the phosphoryl base (phosphorylcholine, phosphorylethanolamine, and others), characteristic of each phospholipid class. Most sphingomyelinases hydrolyze the equivalent bond between ceramide (Cer) and phosphorylcholine in sphingomyelin (SM). From the point of view of their enzyme activities, phospholipases and lipases in general are rather unique among enzymes in that their substrates and end products do not occur freely in solution but are instead found to make up part of the cell membrane.Early work from our laboratory ( 1 ) had shown that phospholipid hydrolysis catalyzed by PLC from Bacillus cereus was able to induce aggregation and fusion of large unilamellar vesicles (LUV). That work was confi rmed by other studies ( 2 ) and was later extended to other PLCs ( 3-7 ). More recently, Montes et al. ( 8 ) described the leakagefree membrane fusion induced by the hydrolytic activity of Abstract The binding and early stages of activity of a phospholipase C/sphingomyelinase from Pseudomonas aeruginosa on giant unilamellar vesicles (GUV) have been monitored using fl uorescence confocal microscopy. Both the lipids and the enzyme were labeled with specifi c fl uorescent markers. GUV consisted of a mixture of phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, and cholesterol in equimolar ratios, to which 5-10 mol% of the enzyme endproduct ceramide and/or diacylglycerol were occasionally added. Morphological examination of the GUV in the presence of enzyme reveals that, although the enzyme diffuses rapidly throughout the observation chamber, detectable enzyme binding appears to be a slow, random process, with new bound-enzyme-containing vesicles appearing for several minutes. Enzyme binding to the vesicles appears to be a cooperative process. After the initial cluster of bound enzyme is detected, further binding and catalytic activity follow rapidly. After the activity has started, the enzyme is not released by repeated washing, suggesting a "scooting" mechanism for the hydrolytic activity. The enzyme preferentially binds the more disordered domains, and, in most cases, the catalytic activity causes the disordering of the other domains. Simultaneously, peanut-or fi gure-eight-shaped vesicles containing two separate lipid domains become spherical. At a further stage of lipid hydrolysis, lipid aggregates are formed and vesicles disintegrate.

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A Cellular Deficiency of Gangliosides Causes Hypersensitivity to Clostridium perfringens Phospholipase C

Cited by 58 publications

References 63 publications

The NanI and NanJ Sialidases of Clostridium perfringens Are Not Essential for Virulence

The NanI and NanJ Sialidases of Clostridium perfringens Are Not Essential for Virulence

Bacterial Sphingomyelinases and Phospholipases as Virulence Factors

Imaging the early stages of phospholipase C/sphingomyelinase activity on vesicles containing coexisting ordered-disordered and gel-fluid domains

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