Adenylate cyclase toxin (ACT) is an important virulence factor secreted by Bordetella pertussis. When a lung infection is underway, ACT interacts with tracheal epithelial cells, inserting itself into cytoplasmic membranes, aiding the adhesion of bacteria to the airway lining. Host phagocytes responding to the site of infection are disabled by cAMP generated by ACT. Encoded by the Bordetella pertussis cyaA gene, adenylate cyclase toxin is single 1706aa polypeptide, with an apparent molecular weight of 220kD. The N-terminal of this protein contains an adenylate cyclase domain which binds to host cell calmodulin and catalyzes unregulated conversion of cellular ATP to cAMP. The C-terminal, receptor binding domain, contains numerous repeat in toxin (RTX) motifs and is responsible for binding to the cellular receptor on the surface of immune cells, translocation to the cell cytosol and creating cation-selective pores in the host cell membrane.
Adenylate cyclase toxin both suppresses and modulates the host immune system contributing to pathogenesis of B. pertussis. This toxin is unusual in its ability to cross mammalian cell membranes and locate within the cytosol. As a cell biology tool, adenylate cyclase has been used to deliver immunogenic epitopes, targeting antigen presenting cells. This protein is the native cell surface antigen but does not exhibit the level of enzyme activity shown by ACT produced in E. coli.
Source:Recombinant protein corresponding to B. pertussis Adenylate Cyclase Toxin.
Molecular Weight:~220kD
Biological Activity:The enzymatic activity of this preparation is greater than 115umoles cAMP/min/mg of protein as determined by a modification of the method of Hewlett et al. The toxicity of this preparation is >9.8umoles cAMP/mg of J774A.1 cell protein/mg toxin as determined by a modification of the method of Hewlett et al.
Storage and Stability:Lyophilized powder may be stored at -20°C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
仕様
Size：50ug
Source Antigen：Recombinant, B. pertussis
Grade：Purified
Purity：~90% (SDS-PAGE)
Form：Supplied as a lyophilized powder in 20mM HEPES, pH 7.5, 8M urea. Reconstitute with 50ul sterile room temperature ddH2O. Handle Gently. Do not vortex.
