Effect of Metal lons on Phospholipase C (Heat-Labile Hemolysin) from Pseudomonas aeruginosa

1997 ◽  
Vol 3 (1) ◽  
pp. 20-26 ◽  
Author(s):  
Shuichi Tawara ◽  
Yoshimi Matsumoto ◽  
Satoru Matsumoto ◽  
Katsuyuki Maki ◽  
Yoshiko Koyama ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phospholipase C ◽  
Heat Labile

Phospholipase C (heat-labile hemolysin) of Pseudomonas aeruginosa: purification and preliminary characterization

1982 ◽  
Vol 152 (1) ◽  
pp. 239-245
Author(s):  
R M Berka ◽  
M L Vasil
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phospholipase C ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Size Exclusion ◽  
Tetradecyltrimethylammonium Bromide ◽  
Heat Labile ◽  
Hydrophobic Moiety ◽  
Culture Supernatants

Phospholipase C (heat-labile hemolysin) was purified from Pseudomonas aeruginosa culture supernatants to near homogeneity by ammonium sulfate precipitation followed by a novel application of DEAE-Sephacel chromatography. Enzymatic activity remained associated with DEAE-Sephacel even in the presence of 1 M NaCl, but was eluted with a linear gradient of 0 to 5% tetradecyltrimethylammonium bromide. Elution from DEAE-Sephacel was also obtained with 2% lysophosphatidylcholine, and to a lesser extent with 2% phosphorylcholine, but not at all with choline. The enzyme was highly active toward phospholipids possessing substituted ammonium groups (e.g., phosphatidycholine, lysophosphatidylcholine, and sphingomyelin); however, it had little if any activity toward phospholipids lacking substituted ammonium groups (e.g., phosphatidylethanolamine, phosphatidylserine, and phosphaditylglycerol). Collectively, these data suggest that phospholipase C from P. aeruginosa exhibits high affinity for substituted ammonium groups, but requires an additional hydrophobic moiety for optimum binding. The specific activity of the purified enzyme preparation increased 1,900-fold compared with that of culture supernatants. The molecular weight of the phospholipase C was estimated to be 78,000 by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Sephacryl S-200 column chromatography and was 76,000 by high-performance size exclusion chromatography. The isoelectric point was 5.5. Amino acid analysis showed that phospholipase C was rich in glycine, serine, threonine, aspartyl, glutamyl, and aromatic amino acids, but was cystine free.

scholarly journals Studies of phospholipase C (heat-labile hemolysin) in Pseudomonas aeruginosa.

1981 ◽  
Vol 34 (3) ◽  
pp. 1071-1074 ◽  
Author(s):  
R M Berka ◽  
G L Gray ◽  
M L Vasil
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phospholipase C ◽  
Heat Labile

FORMATION OF HEMOLYSIN BY STRAINS OF PSEUDOMONAS AERUGINOSA

1966 ◽  
Vol 12 (2) ◽  
pp. 231-241 ◽  
Author(s):  
Robert A. Altenbern
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Particulate Matter ◽  
High Speed ◽  
Heat Labile ◽  
Intracellular Release ◽  
No Release ◽  
Subsequent Exposure

Hemolysin is formed in sonic extracts of cells of Pseudomonas aeruginosa by the action of a heat-labile substance, probably an intracellular "release" enzyme, on a substrate from the disrupted cell. The substrate and most of the hemolysin released can be sedimented by high-speed centrifugation. Hemolysin-negative strains appear to possess no release enzyme but do contain the substrate since addition of particulate matter to extracts of hemolysin-positive cells increases the rate and extent of hemolysin formation. The rate of hemolysin release in sonic extracts is strongly influenced by the concentration of the two reactants, and minor dilution abolishes all activity. There is only a small amount of release enzyme and substrate present in 24-h cells but increasingly greater amounts appear in extracts of 48- and 72-h cells. The hemolysin-forming system is sensitive to heat and is inactivated in 2 min at 100°. Treatment of particulate matter with lysozyme plus EDTA does not reduce the amount of hemolysin released by subsequent exposure of the particles to the release enzyme present in fresh sonic extract.

scholarly journals In vivo studies with two phospholipase C fractions from Pseudomonas aeruginosa.

1987 ◽  
Vol 55 (7) ◽  
pp. 1728-1730 ◽  
Author(s):  
R S Berk ◽  
D Brown ◽  
I Coutinho ◽  
D Meyers
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phospholipase C ◽  
In Vivo Studies

Osmoprotectants and phosphate regulate expression of phospholipase C in Pseudomonas aeruginosa

1992 ◽  
Vol 6 (7) ◽  
pp. 863-871 ◽  
Author(s):  
Virginia D. Shoriridge ◽  
Andrée Lazdunski ◽  
Michael L. Vasil
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phospholipase C

Multiple phospholipid substrates of phospholipase C/sphingomyelinase HR2 from Pseudomonas aeruginosa

2011 ◽  
Vol 164 (1) ◽  
pp. 78-82 ◽  
Author(s):  
David J. López ◽  
M. Isabel Collado ◽  
Maitane Ibarguren ◽  
Adriana I. Vasil ◽  
Michael L. Vasil ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phospholipase C
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