Pertussis Toxin-ATP Complex

A young boy coughing due to pertussis.

Introduction

Pertussis Toxins (PT) is a protein-based exotoxin and major virulence factor produced by the bacterium Bordetella pertussis.^[1] PT causes pertussis, which is also known at whooping cough and is highly contagious bacterial disease. The disease is caused by the bacterium colonizing the respiratory tract where it then establishes an infection. This disease had been characterized by severe coughing that can last up to six weeks and in some countries lasting nearly 100 days.^[2]It has been documented in some cases that PT can cause subconjunctival hemorrhages, rib fractures, hernias, fainting and vertebral artery dissection.

As of 2010, the worldwide incidence of whooping cough has been estimated to 48.5 million cases and nearly 295,000 deaths per year.^[3] With that in mind, whooping cough can affect people of any age; however, before vaccines were available the disease was most common in infants and young children but now children are immunized and the high percentage of cases are seen among adolescents.

structure

The pertussis toxin has been characterized as being an AB toxin meaning that there are 2 subunits: A subunit possesses the enzyme activity and the B possesses it the receptor binding portion. PT in particular is an AB5 toxin consisting of a six-component protein complex, and the multiple subunits of the complex are not identical in composition. With that in mind, this protein is a hexamer containing a catalytic (S1) subunit that is tightly associated with the pentameric cell-binding component (B-oligomer). The S1 component is a single subunit while the B-oligomer is a pentamer composed of four types of subunits: , , two copies of , and .^[1] These subunits are encoded by a ptx genes, which are encoded on a large PT operon that includes additional genes as well such as Pti genes. Together the PT and Pti proteins form the PT secretion complex.

Pertussis Toxin activation

Pertussis Toxin by itself is harmless unless activated. From multiple studies, it has became clear that there is a direct interaction between Adenosine triphosphate (ATP) and pertussis toxin which leads to activation. The direct effect of ATP is to destabilize the interaction between the S1 subunit and the B-oligomer by binding to the B-oligomer. This interaction relaxes the toxin by facilitating the subsequent reduction of a disulphide bond in the S1 subunit. The main interaction that leads to the destabilization is the favorable hydrogen bonding and electrostatic interaction between the triphosphate moiety and five positively charged amino acids:. In contrast, the negatively charged carboxyl terminus of subunit S1 interacts unfavorably with the negative charges of the triphosphate moiety, causing a displacement of the C-terminal of therefore, the repulsion between the triphosphate moiety and the C terminus of subunit S1 forms the mechanism by which the interaction between S1 and the B-Oligomer is destabilized.

Mechanism of pathogenesis

Following the pertussis toxin binding to the cell membrane (B subunit binds to terminal sialic acid residues), the toxin is taken up by an endosome and transported from the plasma membrane via the Golgi apparatus to the endoplasmic reticulum (ER) where finally membrane translocation occurs. The destabilization of PT occurs in the ER prior to membrane translocation. After binding of ATP, cleavage of the single disulphide bond by protein disulphide isomerase (PDI) occurs in subunit S1 and is believed to trigger a conformational change necessary to expose the active site to its substrates. The reducation step takes place after interaction of PT with ATP. After destabilization, the S1 becomes active and catalyzes the ADP-ribosylation of the alpa-subunit of regulatory trimeric G-proteins (Giα). This then prevents Giα from inhibiting adenylate cyclase and leads to an increase in intracellular levels of Cyclic adenosine monophosphate (cAMP). The increase in cAMP affects normal biological signaling and causes severe effects such as hypoglycemia.

Treatment

Early treatment of pertussis is the only effective way to treat the bacterial infection. If treatment for pertussis is started early in the course of illness during the 1 to 2 weeks before severe coughing occurs, the symptoms may be lessened. If the diagnosed to late, antibiotics will not alter the course of the illness since the bacteria is already producing the PT toxin. The preferred antibiotics are Erythromycin, Clarithromycin, and Azithromycin.

The primary method of prevention for pertussis is vaccination. The DTaP vaccine vaccine is used and this vaccine is composed of diphtheria, tetanus, and pertussis. The pertussis component is acellular; this acellular component are selected antigens of pertussis that induces adaptive immunity.

Conclusion

This paper was significant since it gave a clear understanding of the PT activation as well as a better understanding to the pathogenesis of the toxin. The key features of this proposal is that ATP binding signals the arrival of the PT in the endoplasmic reticulum and at the same time triggers dissociation of the holotoxin prior to membrane translocation. Therefore, the dissociation is due to ATP binding destabilization and reduction by protein disulphide isomerase.