The binding and hydrolysis of sphingomyelin by phospholipase C (Bacillus cereus) as shown by 31P NMR

1984 ◽  
Vol 16 (8) ◽  
pp. 931-934 ◽  
Author(s):  
Kjf.ll M. Aalmo ◽  
Jostein Krane ◽  
Clive Little ◽  
Carlyle B. Storm
Keyword(s):  
Bacillus Cereus ◽  
Phospholipase C ◽  
31P Nmr ◽  
Hydrolysis Of

scholarly journals NIH 3T3 cells stably transfected with the gene encoding phosphatidylcholine-hydrolyzing phospholipase C from Bacillus cereus acquire a transformed phenotype.

1994 ◽  
Vol 14 (1) ◽  
pp. 646-654 ◽  
Author(s):  
T Johansen ◽  
G Bjørkøy ◽  
A Overvatn ◽  
M T Diaz-Meco ◽  
T Traavik ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Phospholipase C ◽  
Constitutive Expression ◽  
Soft Agar ◽  
3T3 Cells ◽  
Gene Encoding ◽  
Stable Transfectants ◽  
Nih 3T3 ◽  
Hydrolysis Of ◽  
Nih 3T3 Cells

In order to determine whether chronic elevation of intracellular diacylglycerol levels generated by hydrolysis of phosphatidylcholine (PC) by PC-hydrolyzing phospholipase C (PC-PLC) is oncogenic, we generated stable transfectants of NIH 3T3 cells expressing the gene encoding PC-PLC from Bacillus cereus. We found that constitutive expression of this gene (plc) led to transformation of NIH 3T3 cells as evidenced by anchorage-independent growth in soft agar, formation of transformed foci in tissue culture, and loss of contact inhibition. The plc transfectants displayed increased intracellular levels of diacylglycerol and phosphocholine. Expression of B. cereus PC-PLC was confirmed by immunoperoxidase and immunofluorescence staining with an affinity-purified anti-PC-PLC antibody. The NIH 3T3 clones expressing plc induced DNA synthesis, progressed through the cell cycle in the absence of added mitogens, and showed significant growth in low-concentration serum. Transfection with an antisense plc expression vector led to a loss of PC-PLC expression accompanied by a complete reversion of the transformed phenotype, suggesting that plc expression was required for maintenance of the transformed state. Taken together, our results show that chronic stimulation of PC hydrolysis by an unregulated PC-PLC enzyme is oncogenic to NIH 3T3 cells.

Kinetics of the Hydrolysis of Monodispersed and Micellar Phosphatidylcholines Catalyzed by a Phospholipase C from Bacillus cereus

1991 ◽  
Vol 110 (1) ◽  
pp. 88-95 ◽  
Author(s):  
Kiyoshi Ikeda ◽  
Seiji Inoue ◽  
Chika Amasaki ◽  
Keizo Teshima ◽  
Hiroh Ikezawa
Keyword(s):  
Bacillus Cereus ◽  
Phospholipase C ◽  
Kinetics Of ◽  
Hydrolysis Of

scholarly journals Validation of a simple radiochemical assay measuring hydrolysis of choline-labelled microsomal phosphatidylcholine by phospholipase C. pH-dependence

1976 ◽  
Vol 160 (3) ◽  
pp. 475-479 ◽  
Author(s):  
B R Cater ◽  
P Trivedi ◽  
T Hallinan
Keyword(s):  
Bacillus Cereus ◽  
Phospholipase C ◽  
Clostridium Perfringens ◽  
Ph Dependence ◽  
Selective Hydrolysis ◽  
Ph Range ◽  
Hydrolysis Of

Selective hydrolysis of phosphatidylcholine species, which are selectively radioactively labelled in vivo, does not appear to interfere with a radiochemical assay for hydrolysis of microsomal phosphatidylcholine by C-type phospholipases from Bacillus cereus or Clostridium perfringens. Both phospholipases substantially hydrolysed phosphatidylcholine over the pH range 4.0-10.0.

NIH 3T3 cells stably transfected with the gene encoding phosphatidylcholine-hydrolyzing phospholipase C from Bacillus cereus acquire a transformed phenotype

1994 ◽  
Vol 14 (1) ◽  
pp. 646-654
Author(s):  
T Johansen ◽  
G Bjørkøy ◽  
A Overvatn ◽  
M T Diaz-Meco ◽  
T Traavik ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Phospholipase C ◽  
Constitutive Expression ◽  
Soft Agar ◽  
3T3 Cells ◽  
Gene Encoding ◽  
Stable Transfectants ◽  
Nih 3T3 ◽  
Hydrolysis Of ◽  
Nih 3T3 Cells

In order to determine whether chronic elevation of intracellular diacylglycerol levels generated by hydrolysis of phosphatidylcholine (PC) by PC-hydrolyzing phospholipase C (PC-PLC) is oncogenic, we generated stable transfectants of NIH 3T3 cells expressing the gene encoding PC-PLC from Bacillus cereus. We found that constitutive expression of this gene (plc) led to transformation of NIH 3T3 cells as evidenced by anchorage-independent growth in soft agar, formation of transformed foci in tissue culture, and loss of contact inhibition. The plc transfectants displayed increased intracellular levels of diacylglycerol and phosphocholine. Expression of B. cereus PC-PLC was confirmed by immunoperoxidase and immunofluorescence staining with an affinity-purified anti-PC-PLC antibody. The NIH 3T3 clones expressing plc induced DNA synthesis, progressed through the cell cycle in the absence of added mitogens, and showed significant growth in low-concentration serum. Transfection with an antisense plc expression vector led to a loss of PC-PLC expression accompanied by a complete reversion of the transformed phenotype, suggesting that plc expression was required for maintenance of the transformed state. Taken together, our results show that chronic stimulation of PC hydrolysis by an unregulated PC-PLC enzyme is oncogenic to NIH 3T3 cells.

scholarly journals Asymmetry of lipid organization in cholinergic synaptic vesicle membranes

1983 ◽  
Vol 211 (1) ◽  
pp. 155-162 ◽  
Author(s):  
D M Michaelson ◽  
G Barkai ◽  
Y Barenholz
Keyword(s):  
Bacillus Cereus ◽  
Synaptic Vesicle ◽  
Phospholipase C ◽  
Molar Ratio ◽  
Second Phase ◽  
Outer Leaflet ◽  
Vesicle Membranes ◽  
Vesicular Membrane ◽  
The Asymmetry ◽  
Hydrolysis Of

The lipid composition of purified Torpedo cholinergic synaptic vesicles was determined and their distribution between the inner and outer leaflets of the vesicular membrane was investigated. The vesicles contain cholesterol and phospholipids at a molar ratio of 0.63. The vesicular phospholipids are (mol% of total phospholipids): phosphatidylcholine (40.9); phosphatidylethanolamine (24.6); plasmenylethanolamine (11.5); sphingomyelin (12); phosphatidylserine (7.3); phosphatidylinositol (3.7). The asymmetry of the synaptic vesicle membranes was investigated by two independent approaches: (a) determining accessibility of the amino lipids to the chemical label trinitrobenzenesulphonic acid (TNBS); (b) determining accessibility of the vesicular glycerophospholipids to phospholipase C (Bacillus cereus). TNBS was found to render the vesicles leaky and thus cannot be used reliably to determine the asymmetry of Torpedo synaptic vesicle membranes. Incubation of the vesicles with phospholipase C (Bacillus cereus) results in biphasic hydrolysis of the vesicular glycerophospholipids. About 45% of the phospholipids are hydrolysed in less than 1 min, during which no vesicular acetylcholine is released. In the second phase, the hydrolysis of the phospholipids slows down markedly and is accompanied by loss of all the vesicular acetylcholine. These findings suggest that the lipids hydrolysed during the first phase are those comprising the outer leaflet. Analysis of the results thus obtained indicate that the vesicular membrane is asymmetric: all the phosphatidylinositol, 77% of the phosphatidylethanolamine, 47% of the plasmenylethanolamine and 58% of the phosphatidylcholine were found to reside in the outer leaflet. Since phosphatidylserine is a poor substrate for phospholipase C (B. cereus), its distribution between the two leaflets of the synaptic vesicle membrane is only suggestive.

scholarly journals An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C

2021 ◽  
Author(s):  
Nabangshu Sharma ◽  
Ries J. Langley ◽  
Chatchakorn Eurtivong ◽  
Euphemia Leung ◽  
Ryan Joseph Dixon ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Phospholipase C ◽  
Food Industry ◽  
Maldi Tof ◽  
Hydrolysis Of

The Bacillus cereus phosphatidylcholine-specific phospholipase C (PC-PLCBc) is an enzyme that catalyses the hydrolysis of phosphatidylcholines into phosphocholine and 1,2-diacylglycerols. PC-PLCBc has found applications in both the food industry and...

486. The action of the lecithinase of Bacillus cereus on the globule membrane of milk fat

1952 ◽  
Vol 19 (3) ◽  
pp. 311-315 ◽  
Author(s):  
M. Jean Stone
Keyword(s):  
Bacillus Cereus ◽  
Culture Filtrate ◽  
Microscopic Examination ◽  
Milk Fat ◽  
Marked Effect ◽  
Whole Milk ◽  
Free Choline ◽  
Fat Globule ◽  
Hydrolysis Of ◽  
Qualitative Estimation

An effect simulating broken cream in whole milk was produced when Bacillus cereus was grown in washed cream at 22° C. A similar but less marked effect was obtained when a concentrated culture filtrate of B. cereus was added to washed cream.Microscopic examination of washed cream to which B. cereus or its concentrated filtrate had been added showed that the fat-globule membranes had been broken down. The hydrolysis of lecithin in washed cream in which B. cereus had grown was demonstrated by qualitative estimation of free choline.Since these results could not be reproduced when a non-lecithinase-producing strain of B. cereus or its concentrated culture filtrate was added to washed cream, it appears that the hydrolysis of the lecithin of the fat-globule membrane is at least partly responsible for the formation of broken cream.

scholarly journals Effect of Phospholipase C Hydrolysis of Membrane Phospholipids on Membranous Enzymes

1972 ◽  
Vol 247 (9) ◽  
pp. 2835-2841 ◽  
Author(s):  
Richard D. Mavis ◽  
Robert M. Bell ◽  
P. Roy Vagelos
Keyword(s):  
Phospholipase C ◽  
Membrane Phospholipids ◽  
Hydrolysis Of
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