title: Biological properties of phospholipase C purified from a fleecerot isolate of Pseudomonas aeruginosa.
abstract: A role for one of many exocellular enzymes produced by Pseudomonas aeruginosa--phospholipase C (PLC)--as a prime candidate virulence factor in fleecerot dermatitis has been examined. The addition of Tween 80 in tryptose minimal medium effectively perturbed the membrane system of a field isolate of P. aeruginosa, resulting in increased production and release of a periplasmic enzyme marker, alkaline phosphatase (AP), and also of PLC. PLC activity levels in the culture supernatant were 10- to 15-fold higher in the presence of Tween than in its absence. Apart from AP, the culture medium contained little or no detectable proteolytic enzyme activity, thereby facilitating the partial purification of a haemolytic form of PLC by anion-exchange chromatography. This enzyme, when injected intradermally into the skin of sheep, elicited histopathological lesions virtually identical to those seen in naturally occurring fleecerot. In addition, serum from each of eight sheep afflicted with fleecerot contained high levels of circulating anti-PLC antibody activity when assayed by ELISA. Since these antibodies did not affect the enzymic function of PLC, it is likely that they do not bind to, or are incapable of conformational modification of, the active site.
authors.affiliation: Department of Agriculture, New South Wales, Glenfield, Australia.
authors.forename: J C
authors.initials: JC
authors.lastname: Chin
authors.forename: J E
authors.initials: JE
authors.lastname: Watts
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D003872:Dermatitis / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000254:growth & development / Q000276:immunology; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000382:microbiology*; D012867:Skin / Q000187:drug effects / Q000201:enzymology; D010738:Type C Phospholipases / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000907:Antibodies, Bacterial; D000469:Alkaline Phosphatase; D010738:Type C Phospholipases