Some biological properties of purified staphylococcal haemolysins

Wadström, Torkel; Möllby, R.

doi:10.1016/0041-0101(72)90177-8

Cited by 33 publications

(20 citation statements)

References 38 publications

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“…Recently Mollby and Wadstrom (14,20) reported the isoelectric point of gamma-lysin to be at pH 9.5, but they were unable to confirm that the hemolysin has two components. We have confirmed the separation of gamma-lysin components I and II on hydroxyapatite first reported by Guyonnet et al (8) and shown that the isoelectric points of the two components are extremely close (pH 9.8 and 9.9, respectively).…”

Section: Resultsmentioning

confidence: 97%

Further Characterization of Staphylococcal Gamma-Hemolysin

Taylor

Bernheimer

1974

Infect Immun

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It was confirmed that staphylococcal gamma-hemolysin is composed of two separate proteins (gamma-lysin components I and II) which act synergistically. The molecular weights of the two components, determined by gel filtration, are 29,000 and 26,000, respectively, and their isoelectric points, determined by isoelectric focusing, are at pH 9.8 and 9.9. Both components are susceptible to the action of Pronase and subtilisin. A wide range of lipids, some in minute amounts, are capable of inhibiting the hemolytic activity of gamma-lysin.

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Section: Resultsmentioning

confidence: 97%

Further Characterization of Staphylococcal Gamma-Hemolysin

Taylor

Bernheimer

1974

Infect Immun

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“…The strain was grown in a prereduced proteose peptone medium in a fermentor with controlled pH, temperature and redox-potential (C.-E. Nord, R. Mrllby, & T. Wadstr6m,unpublished). The following methods for analysis of homogeneity of the purified phospholipase were used: (1) acrylamide electrophoresis in different buffer systems [40,41,42]; (2) one precipitin band in immunoelectrophoresis; (3) devoid of any of the 18 different enzyme and toxin activities assayed for [25]. The following methods for analysis of homogeneity of the purified phospholipase were used: (1) acrylamide electrophoresis in different buffer systems [40,41,42]; (2) one precipitin band in immunoelectrophoresis; (3) devoid of any of the 18 different enzyme and toxin activities assayed for [25].…”

Section: Methodsmentioning

confidence: 99%

Effect ofClostridium perfringens phospholipase C (alpha-toxin) on the human diploid fibroblast membrane

1974

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Human diploid embryonic lung fibroblasts were cultivated in Eagle's Minimum Essential Medium and labeled with aH-uridine. The release of soluble radioactive substances into the medium was used as an indicator of damage to the cell membrane. The assay method described is simple, sensitive and rapid and allows quantitative estimation of changes in membrane permeability before any morphological damage is observed microscopically. Crude commercial preparations of phospholipaseC (E.C. 3.1.4.3) (40 rtg/ml) were highly active on the cell membrane but most of the membrane damaging activity was found to be due to contaminating theta-toxin. However, also highly purified phospholipase C caused a membrane damage as measured by release of isotope through the plasma membrane. The release could be increased by including an optimal concentration of calcium ions in the incubation buffer, by treating the cells in a hypotonic medium and by simultaneous treatment with sublytic concentrations of Triton X-100. To our knowledge this is the first report of membrane damage on a live, intact, metabolizing human diploid cell caused by a highly purified phospholipase C. The results are in agreement with a dynamic membrane structure with the polar groups of a part of the phospholipids accessible at the membrane surface.Highly purified phospholipase C (phosphafidylcholine cholinephosphohydrolase, E.C. 3.1.4.3) from Bacillus cereus is nonhaemolytic for erythrocytes of many different species [13,45]. On the other hand, phospholipase C from Clostridium perfringens was shown three decades ago to be haemolytic, dermonecrotic and lethal [20], activities which were later found also in purified preparations [33,37]. However, since this organism also produces other cytolytic proteins such as theta-toxin, it was not clear until recently whether the haemolytic activity was due to contaminating theta-toxin. Commercial, partially purified preparations of this phospholipase C contain this cytolytic factor, as well as about ten other toxins and enzymes [25]. 21 J. Membrane Biol. 16

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“…The S. aureus beta-toxin, P. aeruginosa PLC-H, and C. perfringens alpha-toxin are the most intensively studied in this respect. The beta-toxin shows a marked degree of specificity, being cytotoxic for human thrombocytes but not leukocytes (182). Platelets were lysed rapidly by this toxin (181).…”

Section: Effects Of Phospholipases C On Cells Othermentioning

confidence: 99%

“…The induction of arachidonic acid metabolites by bacterial phospholipases C could account for many of the effects observed with these toxins in vitro and in vivo. Leukotrienes C4 and D4 (LTC4 and LTD4) increase vascular permeability and promote the exudation of fluid into the extravascular space (141,182), and these effects have been observed after the intradermal administration of purified C. perfringens alpha-toxin in the guinea pig (156) and P. aeruginosa PLC-H in the mouse (102). The contraction of the isolated rat aorta and isolated rat ileum could be attributed to the induced production of LTC4 or thromboxane A2 (TXA2) (51).…”

Section: Activation Of Arachidonic Acid Cascadementioning

confidence: 99%