Pertussis
Toxin and Virulence Factors
Pertussis Toxin
Pertussis toxin (PTX) is produced by Bordetella pertussis, the bacterium responsible for whooping cough. Pertussis toxin is a multi-component protein composed of six non-covalently bound subunits ranging in molecular weight from approximately 9 to 28 kDa. The largest subunit, also called the A protomer, is an enzyme responsible for the ADP-ribosyl transferase activity; the A protomer alone will transfer the ADP ribose from NAD+ to α subunits of G proteins of the class Gαi, Gαo or Gαt. The five-subunit binding complex also known as the B oligomer, allows PTX to enter most cells, attaching to glycan residues present on receptor proteins including TLR4 and glycoprotein Ib. After entering the cell via receptor-mediated endocytosis, PTX is transported retrogradely via the endosomal pathway and Golgi complex to the endoplasmic reticulum. The A subunit is released from the toxin and translocates through the membrane of the endoplasmic reticulum where the toxin inactivates the target membrane-bound G proteins. Cell biologists use PTX to eliminate G proteins and thereby identify Gi/o protein-dependent pathways. Since toxin receptor is present in several mammalian cell types, pertussis toxin affects most cultured cells. Additionally, pertussis toxin helps to create animal models of autoimmune disease. For example, pertussis toxin is administered to mice along with myelin antigen to create a mouse model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE). List labs offers native pertussis toxin, isolated from Bordetella pertussis strain 165. In addition, List Labs also produces a genetically inactive form of pertussis toxin as well as several of the virulence factors associated with Bordetella pertussis infections. Pertussis Toxin preparations are highly purified, containing all five subunits. Preparations are tested to assure minimal contamination with endotoxin, adenylate cyclase and other virulence factors. Each lot is tested for activity in CHO cells. Native pertussis toxin is supplied in three formulations described below:
•Product #179 Pertussis
toxin from B. pertussis strain
165 in glycerol is provided frozen in a glycerol-containing Tris buffer.
It will be shipped on ice.
•Product #180 Pertussis
toxin from B. pertussis strain
165, lyophilized in buffer is offered freeze-dried in phosphate buffer.
•Product #181 Pertussis
toxin from B. pertussis strain
165, lyophilized salt-free is freeze-dried in water and may be reconstituted
with water or the buffer of choice.
Pertussis Toxin Mutant
List Labs sells a genetically inactivated form of pertussis toxin, Pertussis Toxin Mutant (PTMUT). This protein may be used as an antigen and as a carrier protein for other antigens. Pertussis mutant protein is isolated from the TY-178 strain of Bordetella bronchiseptica. Site-directed mutagenesis was used to replace amino acid residues in the enzyme active site. Although diminished, this mutant retains some of the activity of the native toxin.
•Product #184 Pertussis
toxin mutant (R9K, E129A) is provided lyophilized in phosphate buffer and an
accompanying C of A describes the CHO cell toxicity.
Products
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