Purified macrophage cationic proteins were used in functional assays to determine their interactions with the outer membrane and lipopolysaccharide of Pseudomonas aeruginosa. A fluorescent derivative of polymyxin B (dansyl-polymyxin) was found to bind to saturation to purified lipopolysaocharide, with similar affinity for the aminoglycoside supersensitive strain H215 and wild type strain H103 lipopolysaocharide. MCP-1 could displace more dansyl-polymyxin bound to the lipopolysaocharide of both strains, and bound with greater affinity than MCP-2. When whole cells were used, MCPs also displaced bound dansyl-polymyxin. Effects on the outer membrane of whole cells were examined by determining the initial rate of uptake of the hydrophobic fluorescent probe 1-N-phenylnaphthylamine. Uptake was enhanced in the presence of MCPs, indicating permeabilization of the outer membrane. MCP-1 caused maximal uptake of the probe at 40 µg/ml, MCP-2 at 70 µg/ml, and crude extract at only 20 µg/ml. Uptake of the probe was found to be enhanced at add pH, with maximal uptake occurring with only 7.5 µg/ml MCP-1 at pH 6.5. The data suggested that MCPs act to permeabilize the outer membranes of P. aeruginosa in a manner analagous to that defined for other polycationic agents. / Science, Faculty of / Microbiology and Immunology, Department of / Graduate
| Identifer | oai:union.ndltd.org:UBC/oai:circle.library.ubc.ca:2429/26531 |
| Date | January 1987 |
| Creators | Sawyer, Janet Gail |
| Publisher | University of British Columbia |
| Source Sets | University of British Columbia |
| Language | English |
| Detected Language | English |
| Type | Text, Thesis/Dissertation |
| Rights | For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use. |
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