Site-directed mutagenesis of the Pseudomonas aeruginosa cytotoxin for probing toxic activity.
The location of amino acids with direct influence on toxic activity in the pore-forming cytotoxin from Pseudomonas aeruginosa was tested by site-directed mutagenesis. Mutant fragments obtained by the polymerase-chain reaction were subcloned into a cytotoxin gene-bearing plasmid to minimize the possibility of amplification error. Our data suggest an important role of cysteines for toxic activity of cytotoxin. Furthermore, domain-facilitating-binding to plasma membranes is located on the C-terminal side of Cys215.
Xiong
G
G
Institut für Pharmakologie und Toxikologie, Universität Giessen, Federal Republic of Germany.
Lutz
F
F
D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D003603:Cytotoxins / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D005814:Genes, Viral; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D016133:Polymerase Chain Reaction; D011550:Pseudomonas aeruginosa / Q000378:metabolism*
D003603:Cytotoxins