描述
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. These subunits are designated as S1, S2, S3, S4 and S5 and occur in native pertussis toxin
in a ratio of 1:1:1:2:1, where the subunit S4 is present in two copies.1, 2
The largest subunit S1, also called the A
protomer, is responsible for the ADP-ribosyltransferase 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 crystal structure of PTX reveals a pyramid-like shape
with the A protomer situated on top of the S5 subunit, which rests on two dimers, S2-S4 and S3-S4.3 Together the five
subunit platform is called the B oligomer, and under certain conditions PTX dissociates into just two parts, the enzymatic
A protomer and the five subunit binding complex, the B oligomer. This B oligomer allows PTX to enter most cells,
attaching to glycan residues present on receptor proteins including TLR44 and glycoprotein Ib.5 After entering the cell
via receptor-mediated endocytosis, PTX is transported retrogradely via the endosomal pathway and Golgi complex to
the endoplasmic reticulum. A protomer is released from the toxin and translocates through the membrane of the
endoplasmic reticulum where the toxin inactivates the target membrane-bound G proteins.6, 7
In mammalian cells, G proteins serve to keep cellular adenylate cyclase in check. Within these cells, pertussis toxin
catalyzes the breakdown of cellular NAD+
, transferring the ADP-ribose moiety to alpha subunits of G proteins of the
heterotrimeric Gi/o protein subfamily. The covalent modification of G proteins disrupts signaling pathways where G
protein binding is part of the chain of events.8,9
When a signaling molecule binds to a G protein-coupled receptor on the
surface of a pertussis-intoxicated cell, G protein is not available to pass on the message to intracellular effectors. Thus,
ADP-ribosylated G proteins can no longer inhibit the cellular adenylate cyclase, allowing production of cyclic AMP,
without restraint. Since cAMP is a key signaling molecule, accumulation of cAMP as a result of PTX intoxication
stimulates cAMP mediated pathways. Many of the effects of pertussis toxin are due to this process.
Also, since G proteins are present in several mammalian cell types, pertussis toxin affects most cultured cells. In cell
biology, pertussis toxin is useful in eliminating and thereby identifying Gi/o protein-dependent pathways. Gi/o proteindependent effects of pertussis toxin are numerous and include lymphocytosis,10,11 histamine sensitization12 and insulin
secretion which is reviewed by Straub et al.
13
Additionally, pertussis toxin appears to act through a phosphokinase C
pathway to increase the permeability of the blood-brain barrier leading to neurological effects.14
Some of the effects of PTX are due to the binding of the toxin to the cell, not to the enzymatic inactivation of G
proteins. These effects are often shown to occur with PTX, or with either toxoid or genetically engineered toxin with an
altered enzyme activity. Studies indicate that mutant pertussis toxin possesses adjuvant properties with the ability to
encourage both local and systemic responses, to promote T helper cell responses to co-administered antigens and to
favor the production of Th1/Th17 cells, important in mediating host immunity to infectious pathogens.15
PTX binds to
the cell receptor, TLR4 which activates Rac and subsequently causes various effects depending on the type of cell
treated.4 The toxin or binding oligomer induces dendritic cell maturation in a TLR4-dependent manner.16 Effects of PTX
intoxication, both dependent and independent of G proteins are reviewed by Locht et al7 and Mangmool et al.17
Pertussis toxin has been variously referred to in literature as lymphocytosis-promoting factor, islet-activating protein,
histamine-sensitizing factor, hemagglutinin and pertussigen. The toxin is useful in creating 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).18, 19
Pertussis toxin from List Biological Laboratories is isolated from Bordetella pertussis strain 165. This preparation is highly
purified and contains all five subunits. It is tested to assure minimal adenylate cyclase activity in the presence of
calmodulin.
20,21 Each lot is tested for activity in the CHO cell assay as described by Hewlett et al.22
©2012, LBL, Inc., Rev. 09/2017
List Labs produces Pertussis Toxin Mutant R9K, E129A, a genetically inactivated mutant of pertussis toxin, which has a
modified sequence encoding the enzyme subunit. Virulence of this pertussis mutant is reduced relative to that found with
the wild type.
23 The pertussis mutant protein is isolated from the TY-178 strain of Bordetella bronchiseptica which
contains a genetically modified sequence encoding the S1 subunit. Genetically inactivated toxin may be used as an
antigen or as a carrier to promote an immune response.
Native pertussis toxin is supplied in three formulations (List Labs Prod. #179, 180 and 181), in glycerol, lyophilized, and
lyophilized salt-free. Pertussis Toxin Mutant (Prod. #184) is supplied lyophilized. A detailed lot analysis documenting
purity and biological activity plus complete instructions on reconstitution and storage accompany each shipment.
These products are intended for research purposes only and are not for use in humans or as diagnostic agents.
For further information, please contact List Biological Laboratories, Inc.
Product No. Description Size
179A,B Pertussis Toxin, in Glycerol 50 μg, 200 µg
180 Pertussis Toxin, Lyophilized in Buffer 50 μg
181 Pertussis Toxin, Lyophilized, Salt-Free 50 μg
184 Pertussis Toxin Mutant 50 µg
Related Products
Filamentous Hemagglutinin (FHA) from B. pertussis, Product #170
Fimbriae 2/3 from B. pertussis, Product #186
Pertactin from B. pertussis (69 kDa Protein), Product #187
Adenylate Cyclase Toxin, Recombinant from B. pertussis, Frozen Liquid, Product #188L
Adenylate Cyclase, Recombinant, Reduced Endotoxin, Frozen Liquid, Product #197L
Adenylate Cyclase Toxoid, Recombinant, Frozen Liquid, Product #198L
HPT™ LPS, highly purified from B. pertussis 165, Product #400
See how others have used List Labs’ products on our citations page: https://www.listlabs.com/citations
References
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protein, pertussis toxin, in conformity with the A-B model. Biochemistry. 1982; 21:5516–5522. PMID:6293544
2. Moss J, Manganiello VC, and Vaughan M. Hydrolysis of nicotinamide adenine dinucleotide by choleragen and its A
protomer: possible role in the activation of adenylate cyclase. Proc Natl Acad Sci USA. 1976; 73:4424-4427.
PMCID: PMC431483
3. Stein PE, Boodhoo A, Armstrong GD, Cockle SA, Klein MH, Read RJ. The crystal structure of pertussis toxin.
Structure. 1994; 2:45–57. PMID:7634099
4. Nishida, M, Suda R, Nagamatsu Y, Tanabe S, Onohara N, Nakaya M, Kanaho Y, Shibata T, Uchida K, Sumimoto,
Sato Y, Kurose H. Pertussis toxin up-regulates angiotensin type 1 receptors through Toll-like receptor 4-mediated
Rac activation. J Biol Chem. 2010; 285:15268–15277. PMCID: PMC2865339
5. Sindt KA, Hewlett EL, Redpath GT, Rappuoli R, Gray LS, Vandenberg SR. Pertussis toxin activates platelets
through an interaction with platelet glycoprotein Ib. Infect Immun. 1994; 62:3108–3114. PMCID: PMC302934
6. Plaut RD and Carbonetti NH. Retrograde transport of pertussis toxin in the mammalian cell. Cell Microbiol. 2008;
10:1130-9. PMID:18201245
7. Locht C, Coutte L, Mielcarek N. The ins and outs of pertussis toxin. FEBS Journal. 2011; 278:4668–4682.
PMID:21740523
8. Katada T, Oinuma M, Ui M. Two guanine nucleotide-binding proteins in rat brain serving as the specific substrate of
islet-activating protein, pertussis toxin. Interaction of the alpha-subunits with beta gamma-subunits in development
of their biological activities. J Biol Chem. 1986; 261:8182-8191. PMID:3087970
9. Kurose H, Katada T, Amano T, Ui M. Specific uncoupling by islet-activating protein, pertussis toxin, of negative
signal transduction via alpha-adrenergic, cholinergic, and opiate receptors in neuroblastoma x glioma hybrid cells. J
Biol Chem. 1983; 258:4870–4875. PMID:6300102
10. Sato Y, Arai H and Suzuki K. Leukocytosis-promoting factor of Bordetella pertussis. 3. Its identity with protective
antigen. Infect Immunity 1974; 9:801-810. PMCID: PMC414888
11. Morse SI and Morse JH. Isolation and properties of the leukocytosis- and lymphocytosis- promoting factor of
Bordetella pertussis. J Exp Med. 1976; 143:1483-1502. PMCID: PMC2190226
12. Muñoz JJ and Bergman RK. Histamine-sensitizing factors from microbial agents, with special reference to
Bordetella pertussis. Bacteriol Rev. 1968; 32:103-126. PMCID: PMC378300
13. Straub SG and Sharp GWG. Evolving insights regarding mechanisms for the inhibition of insulin release by
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PMID:22492651
14. Brückener KE, el Bayâ A, Galla H-J and Schmidt MA. Permeabilization in a cerebral endothelial barrier model by
pertussis toxin involves the PKC effector pathway and is abolished by elevated levels of cAMP. Journal of Cell
Science. 2003; 116:1837-1846. PMID:12665564
15. Nasso M, Fedele G, Spensieri F, Palazzo R, Costantino P, Rappuoli R, Ausiello CM. Genetically detoxified
pertussis toxin induces Th1/Th17 immune response through MAPKs and IL-10-dependent mechanisms. J
Immunol. 2009; 183:1892–1899. PMID:19596995
16. Wang ZY, Yang D, Chen Q, Leifer CA, Segal DM, Su SB, Caspi RR, Howard ZO, Oppenheim JJ. Induction of
dendritic cell maturation by pertussis toxin and its B subunit differentially initiate Toll-like receptor 4-dependent
signal transduction pathways. Exp Hematol. 2006; 34:1115–1124. PMID:16863919
17. Mangmool, S and Kurose, H. G(i/o) protein-dependent and -independent actions of Pertussis Toxin (PTX). Toxins.
2011; 3:884-899. PMCID: PMC3202852
18. Munoz JJ, Bernard CC, Mackay IR. Elicitation of experimental allergic encephalomyelitis (EAE) in mice with the
aid of pertussigen. Cell Immunol. 1984; 83:92-100. PMID:6607126
19. Chen X, Winkler-Pickett RT, Carbonetti NH, Ortaldo JR, Oppenheim JJ & Howard OM. Pertussis toxin as an
adjuvant suppresses the number and function of CD4+CD25+ T regulatory cells. Eur J Immunol. 2006; 36:6716–
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