Recombinant Bordetella pertussis Adenylate Cyclase Toxin

Nature

Recombinant

Tag/Conjugate

Unconjugated

Molecular Weight

~200kDa

Endotoxin

The endotoxin content, determined using a kinetic chromogenic LAL assay, is <130 EU/mg.

Alternative Names

Bordetella pertussis Adenylate Cyclase Toxin

Purity

When examined on a 3-8% SDS-polyacrylamide gel, this protein migrates as a single major band with an apparent molecular weight of 200kDa. Densitometric analysis indicates that the purity is ~90%.

Format

Frozen Liquid

Buffer

0.05M Tris, 8M Urea, 0.002M EDTA, pH8.0

Preservative

None

Storage

Aliquots, sized for single use, may be frozen at -20°C to avoid repeated thawing and freezing of samples. Frozen ACT is stable for years, and upon renaturation, if sufficiently diluted out of urea, it efficiently recovers the activity of binding and penetrating cells.

Antigen Description

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 1706 amino acid polypeptide, with an apparent molecular weight of 220 kDa. 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.