Some Characteristics of Phospholipase C from <i>Bacillus cereus</i>

Otnaess, A B; Little, C. H. A.; Sletten, Knut; Wallin, Reidar; Johnsen, S.; Flengsrud, Ragnar; Prydz, Hans

doi:10.1111/j.1432-1033.1977.tb11828.x

Cited by 94 publications

(34 citation statements)

References 61 publications

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“…5). In both preparations, a phospholipase C from B. cereus, which hydrolyzes glycerol phospholipids such as lecithin, phosphatidylethanolamine, and phosphatidylserine, but not phosphatidylinositol (Otnaess et al, 1977), did not release axonin-1. In the presence of ZnClz, which is known to inhibit PtdIns-PLC (Ikezawa and Taguchi, 1981), axonin-1 was not released, indicating that PtdIns-PLC was indeed responsible for axonin-I release and arguing against release by proteolytic cleavage due to proteases contained in the membranes or included as contaminations of the enzyme preparation.…”

Section: Axonin-1 Is Membranebound By a Glycosyl-ptdins Anchormentioning

confidence: 93%

The axonally secreted cell adhesion molecule, axonin‐1

Zuellig

Rader

Schroeder

et al. 1992

European Journal of Biochemistry

120

111

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Axonin-I is an axon-associated cell adhesion molecule (AxCAM) of the chicken, which promotes neurite outgrowth by interaction with the AxCAM Ll(G4) of the neuritic membrane. Here we report the cloning and sequence determination of a cDNA encoding axonin-I, Peptides generated by enzymatic cleavage showed similarity to the AxCAM F11. Degenerated polymerase chain reaction (PCR) primers were designed and an axonin-I fragment was amplified from mRNA of embryonic retina. Screening of a cDNA library from embryonic brain resulted in the isolation of a 4.0-kb cDNA insert with an open reading frame of 3108 nucleotides. The deduced polypeptide of 1036 amino acids includes a putative hydrophobic N-terminal signal sequence of 23 or 25 amino acids and a C-terminal hydrophobic sequence of 29 amino acids which is suggestive of sequences serving as signal for the attachment of a glycosyl-phosphatidylinositol (glycosyl-PtdIns) anchor. The putative mature form of axonin-1 comprises six immunoglobulin-like repeats, followed by four fibronectin-type I11 repeats.Axonin-1 exhibits 75% amino acid identity with the AxCAM TAG-1 of the rat, suggesting that it is the chicken homologue of TAG-1. Like TAG-1, axonin-I is glycosyl-PtdIns-anchored to the neuronal membrane; in contrast to TAG-1, it does not exhibit an Arg-Gly-Asp sequence.The interconnection of neurons by processes forming synaptic contacts is one of the crucial developmental stages of neurogenesis. In order to span long distances, bundles of axons are formed in a process which is thought to be driven by the tendency of growing axons to adhere specifically to and elongate along preexisting axons. The consistency with which

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Section: Axonin-1 Is Membranebound By a Glycosyl-ptdins Anchormentioning

confidence: 93%

The axonally secreted cell adhesion molecule, axonin‐1

Zuellig

Rader

Schroeder

et al. 1992

European Journal of Biochemistry

120

111

View full text Add to dashboard Cite

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“…This occurs during lipid depletion due to exogenous phospholipase action as well as during accumulation of phospholipids due to biosynthesis in the cytoplasmic layer of the membrane. The latter process as well as a subsequent fast transbilayer movement of the newly synthesized phosphatidylethanolamine was shown by Rothman and Kennedy studying B. megaterium membranes [16]. Also in erythrocytes newly synthesized phospholipid is translocated readily to the outer membrane layer [18].…”

Section: Discussionmentioning

confidence: 92%

“…The residual phospholipids, cardiolipin and 2'-glucosaminylphosphatidylglycerol were found to be largely resistant against the enzyme. This may be due to the known low activity of phospholipase C towards these lipids [2,16] or to a protected position of the lipids in the membrane arising from interaction of lipids with membrane proteins. The second type of control experiments should demonstrate that the reagents do not have access to the lipids in the cytoplasmic layer of the membrane.…”

Section: Discussionmentioning

confidence: 99%

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Localization of Phospholipids in the Membrane of Bacillus megaterium

Demant¹,

Kamp²,

Deenen³

1979

European Journal of Biochemistry

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Phospholipase C treatment of Bacillus megaterium protoplasts at 37 "C removed up to 63% of the total phospholipid without disrupting the membrane. More than 95% of the total phosphatidylethanolamine and 3'-glucosaminylphosphatidylglycerol, 80 % of the phosphatidylglycerol and about 20 % of the cardiolipin and 2'-glucosaminylphosphatidylglycerol were hydrolyzed.Similar results were obtained with isolated membranes and dispersions of lipids extracted from the membranes, demonstrating that these data do reflect, at least in part, the substrate specificity of phospholipase C and not necessarily the distribution of phospholipids over the two membrane layers.When protoplasts and isolated membranes were incubated with excess phospholipase C at temperatures inbetween 5 "C and 25 "C a different hydrolysis pattern was obtained and it was concluded that 50% of the phosphatidylethanolamine and 70% of the 3'-glucosaminylphosphatidylglycerol are located in the outer membrane layer, the residual being in the cytoplasmic layer. At least 40% of the phosphatidylglycerol was found to be present also in the outer layer. The data do not allow conclusions about the localization of cardiolipin and 2'-glucosaminylphosphatidylglycerol. Trinitrobenzenesulfonic acid reacted with 50% of the phosphatidylethanolamine in protoplasts and with nearly 100% of this lipid in isolated membranes at various temperatures and concentrations, thus confirming the symmetric localization of this lipid. The data furthermore indicate that a temperature-dependent transbilayer movement of phospholipids is induced by phospholipase C treatment of the Bacillus megaterium protoplasts.

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“…The most important classes of phospholipases that have been shown to play a significant role in bacterial pathogenesis are phospholipase C and phospholipase D (31). Phospholipase C has been demonstrated to be an important virulence factor in an increasing number of bacteria, including Clostridium perfringens (22,23), Bacillus cereus (25), the intracellular pathogen Listeria monocytogenes (32,37), the extracellular pathogen Pseudomonas aeruginosa (24,34), and other bacteria (20,35).…”

Section: No Hemolytic Activity Was Detected In M Smegmatis M Gordomentioning

confidence: 99%

Detection of Phospholipase C in Nontuberculous Mycobacteria and Its Possible Role in Hemolytic Activity

et al. 2001

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Phospholipase C plays a key role in the pathogenesis of several bacterial infections, for example, those caused by Clostridium perfringens and Listeria monocytogenes. Previous studies have reported multiple copies of plc genes homologous to Pseudomonas aeruginosa plcH and plcN genes encoding the hemolytic and nonhemolytic phospholipase C enzymes in the genomes of Mycobacterium tuberculosis, M. marinum, M. bovis, and M. ulcerans. In this study we analyzed the possible relationship between phospholipase C and hemolytic activity in 21 strains of nontuberculous mycobacteria representing nine different species. Detection of phospholipase C enzymatic activity was carried out using thin-layer chromatography to detect diglycerides in the hydrolysates of radiolabeled phosphatidylcholine. DNA sequences of M. kansasii and M. marinum homologous to the genes encoding phospholipase C from M. tuberculosis and M. ulcerans were identified by DNA-DNA hybridization and sequencing. Finally, we developed a direct and simple assay to detect mycobacterial hemolytic activity. This assay is based on a modified blood agar medium that allows the growth and expression of hemolysis of slow-growing mycobacteria. Hemolytic activity was detected in M. avium, M. intracellulare, M. ulcerans, M. marinum, M. tuberculosis, and M. kansasii mycobacteria with phospholipase C activity, but not in M. fortuitum. No hemolytic activity was detected in M. smegmatis, M. gordonae, and M. vaccae. Whether or not phospholipase C enzyme plays a role in the pathogenesis of nontuberculous mycobacterial diseases needs further investigation.Phospholipases have a wide spectrum of in vivo and in vitro effects, ranging from minor alterations in cell membrane composition and function to lethality. The most important classes of phospholipases that have been shown to play a significant role in bacterial pathogenesis are phospholipase C and phospholipase D (31). Phospholipase C has been demonstrated to be an important virulence factor in an increasing number of bacteria, including Clostridium perfringens (22, 23), Bacillus cereus (25), the intracellular pathogen Listeria monocytogenes (32, 37), the extracellular pathogen Pseudomonas aeruginosa (24, 34), and other bacteria (20,35).Phospholipase genes homologous to the hemolytic plcH and nonhemolytic plcN genes from P. aeruginosa have been described in Mycobacterium tuberculosis, M. bovis, M. marinum, and recently in M. ulcerans (3,11,16). Fusion of mpcA or mpcB plc genes of M. tuberculosis encoding glutathione S-transferase produced beta-hemolytic activity when expressed in Escherichia coli (19), and phospholipase C sphingomyelinase activity was detected in cell extracts from M. smegmatis harboring recombinant mpcA or mpcB genes (16).Nontuberculous mycobacteria are free-living saprophytes that have been detected and isolated from a variety of environmental sources, including water, soil, and dust (6), and some of these mycobacteria are opportunistic pathogens.Nontuberculous mycobacteria, including M. kansasii and M. avium-M....

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Some Characteristics of Phospholipase C from Bacillus cereus

Cited by 94 publications

References 61 publications

The axonally secreted cell adhesion molecule, axonin‐1

The axonally secreted cell adhesion molecule, axonin‐1

Localization of Phospholipids in the Membrane of Bacillus megaterium

Detection of Phospholipase C in Nontuberculous Mycobacteria and Its Possible Role in Hemolytic Activity

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