Bordetella pertussis adenylate cyclase toxin as a tool to analyze molecular interactions in a bacterial two-hybrid system.
Bordetella pertussis secretes a calmodulin-activated adenylate cyclase toxin (CyaA) that is able to enter into eukaryotic cells. We took advantage of the modular structure of the catalytic domain of CyaA to design a genetic system that can detect protein-protein interactions in Escherichia coli. This bacterial two-hybrid system is based on the functional complementation between two complementary fragments, T25 and T18, of the catalytic domain of CyaA, in an E. coli cya strain. This bacterial two-hybrid system could find applications in the studies of structure/function relationships of proteins, in functional analysis of genomes, in high-throughput screening of interacting ligands and in design of new therapeutic agents.
Karimova
G
G
Unité de Biochimie Cellulaire, CNRS URA 2185, Institut Pasteur, Paris, France.
Ullmann
A
A
Ladant
D
D
D037361:Adenylate Cyclase Toxin; D001886:Bordetella pertussis / Q000472:pathogenicity*; D020134:Catalytic Domain; D004926:Escherichia coli / Q000235:genetics*; D006824:Hybridization, Genetic; D010446:Peptide Fragments / Q000235:genetics*; D013329:Structure-Activity Relationship; D010566:Virulence Factors, Bordetella / Q000737:chemistry / Q000235:genetics* / Q000633:toxicity
D037361:Adenylate Cyclase Toxin; D010446:Peptide Fragments; D010566:Virulence Factors, Bordetella