Expression of pertussis toxin subunit S4 as an intracytoplasmic protein in Bacillus subtilis (2023)

Cited by (12)

  • Production of Recombinant Proteins in Bacillus subtilis

    2007, Advances in Applied Microbiology

    The classical way of protein purification starting from a large number of cells producing the protein using its authentic expression signals has been replaced by recombinant technology, where the protein of interest is overproduced in a regulated way. This often means that the recombinant gene is fused to a controllable promoter, which is activated by addition of an inducer to initiate transcription. In most cases, the inducer is a small molecule that is either taken up by the cells or diffuses through the cytoplasmic membrane such as isopropyl thiogalactose (IPTG), xylose, glycine. Overproduction of recombinant proteins is always a two-step process starting with a growth regimen to obtain a high cell density followed by the expression phase. First, cells are grown into the appropriate medium into the appropriate growth phase (with bacteria normally into the mid- or late-exponential growth phase) and then cells will be induced.

  • Assessment of non-protein impurities in potential vaccine proteins produced by Bacillus subtilis

    1993, Vaccine

    The levels of non-protein impurities at different stages of purification of model vaccine proteins produced by Bacillus subtilis were assessed with special emphasis on peptidoglycan-wall teichoic acid and lipoteichoic acid. Intracytoplasmically produced proteins were purified by disrupting the lysozyme protoplasts using osmotic shock, depositing the inclusion bodies by low-speed centrifugation, and washing them with detergent. By this procedure most of the cell envelope-derived impurities could be removed. The final product contained less than 1% (w/w) of neutral sugars, fatty acids, phosphate, hexosamine, diaminopimelic acid and glycerol. A secreted protein was purified from the culture supernatant by successive ion-exchange and adsorption chromatography. The cell envelope-derived impurities were efficiently removed by the cation-exchanger, and the final product contained only minute amounts of non-protein components. The amounts of non-protein components such as peptidoglycan and lipoteichoic acid in proteins produced in either mode were shown to be negligible in relation to their potentially harmful biological effects.

  • High level production of Echerichia coli outer membrane proteins OmpA and OmpF intracellularly in Bacillus subtilis

    1991, FEMS Microbiology Letters

    A high yield of Escherichia coli outer membrane proteins OmpA (about 200 mg/l) and OmpF (about 100 mg/l) was obtained in Bacillus subtilis when produced intracellularly. The yield was more than 100-fold higher than the yield of these proteins by a similar vector containing the complete signal sequence of α-amylase of B. amyloliquefaciens. Both proteins isolated after breakage of the B. subtilis cells by low-speed centrifugation were about 70% pure and could be solubilized by Sarkosyl, SDS and guanidine hydrochloride.

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What is the mechanism of action of pertussis toxin? ›

Pertussis toxin (PT) utilizes a retrograde transport pathway through the Golgi complex and endoplasmic reticulum (ER) to gain entry to the cell cytosol. ATP binding to PT B oligomer (PTB) in the ER promotes holotoxin dissociation for further transport of the active subunit.

How does pertussis toxin affect G-proteins? ›

Pertussis toxin (PTX) has been widely used as a reagent to characterize the involvement of heterotrimeric G-proteins in signalling. This toxin catalyses the ADP-ribosylation of specific G-protein alpha subunits of the Gi family, and this modification prevents the occurrence of the receptor-G-protein interaction.

What is the toxin in pertussis? ›

Pertussis toxin (PT) is a protein-based AB5-type exotoxin produced by the bacterium Bordetella pertussis, which causes whooping cough. PT is involved in the colonization of the respiratory tract and the establishment of infection.

What is the S1 subunit of pertussis toxin? ›

Pertussis toxin (PT) is an AB-type protein toxin that consists of an enzymatic A moiety and a cell-binding B moiety (reviewed in (1, 2)). PT A (the S1 subunit) activates certain Gα proteins by an ADP-ribosylation reaction that utilizes NAD as a donor molecule.

What type of enzyme activity does pertussis toxin have? ›


Part A is an enzymatically active subunit (S1) that has ADP ribosyltransferase activity on certain members of the G-protein family located on mammalian cells that are involved in transduction of receptor signals on cell membranes.

What is the cell signaling pathway involved in pertussis? ›

Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells.

What are the three toxins produced by Bordetella pertussis that cause damage to the host? ›

Bordetella pertussis produces several toxins that affect host-pathogen interactions. Of these, the major toxins that contribute to pertussis infection and disease are pertussis toxin, adenylate cyclase toxin-hemolysin and tracheal cytotoxin.

Which Gram-negative bacilli causes whooping cough? ›

Bordetella pertussis

pertussis is a fastidious, gram-negative bacterium requiring special media for isolation. B. pertussis produces multiple antigenic and biologically active products including: Pertussis toxin.

What are pertussis toxin sensitive G proteins? ›

Pertussis toxin (PTX)-sensitive GTP-binding proteins (G proteins) are essential intermediaries subserving neuronal signal transduction pathways that regulate excitation-secretion coupling.

What vaccines are pertussis toxin? ›

Acellular pertussis vaccine is combined with tetanus toxoid and diphtheria toxoid as DTaP (Infanrix and Daptacel) or Tdap (Boostrix and Adacel). DTaP and Tdap contain the same pertussis components, but Tdap contains a reduced quantity of some pertussis antigens and diphtheria toxoid.

How do you treat pertussis toxin? ›

Antibiotic treatment is available and important to prevent further transmission. However, antibiotics only reduce symptoms if administered in early stages, which rarely occurs due to a late diagnosis. Thus, no causative treatments against symptoms of whooping cough are currently available.

Is pertussis toxin an antigen? ›

Clearly, PT is a key protective antigen in pertussis vaccines, despite the lack of a full understanding of its mechanism of action. PT is responsible for specific pertussis symptoms, but by itself does not recapitulate the full spectrum of disease.

What is the subunit of pertussis? ›

Pertussis toxin is a member of the AB5 family of toxins and is composed of five subunits (S1 to S5) present in a 1:1:1:2:1 ratio. Secretion is a complex process. Each subunit has a secretion signal that mediates transport to the periplasm, where processing and assembly occur.

What are the three types of pertussis? ›

There are three recognized stages of the disease: catarrhal, paroxysmal, and convalescent. The incubation period for Pertussis is 7 to 10 days.

Which receptor is sensitive to pertussis toxin? ›

Stimulation of GABAB1a+2 receptors significantly increased current density, and this was sensitive to PTx.

Is pertussis a culture or PCR? ›

Polymerase chain reaction (PCR) is an important tool for timely diagnosis of pertussis and is increasingly available to clinicians. PCR is a molecular technique used to detect DNA sequences of the Bordetella pertussis bacterium and unlike culture, does not require viable (live) bacteria present in the specimen.

Where is pertussis bacteria found? ›

It is a highly infectious bacterial disease involving the respiratory tract. It is caused by a bacterium (Bordetella pertussis or Bordetella parapertussis ) that is found in the mouth, nose and throat of an infected person.

What bacteria is pertussis cough? ›

Whooping cough, also known as pertussis, is a very contagious respiratory illness caused by a type of bacteria called Bordetella pertussis. The disease is only found in humans.

How does the immune system response to pertussis? ›

With time, the body's immune system develops antibodies which are specialised immune proteins that recognise that whooping cough bacteria are foreign. The antibodies stick to the bacteria and help the body's immune cells to respond to fight the infection.

What cells does pertussis infect? ›

Bordetella pertussis, the causative agent of whooping cough, is internalized by several cell types, including epithelial cells, monocytes, and neutrophils. Although its ability to survive intracellularly is still debated, it has been proven that cell-mediated immunity (CMI) plays a pivotal role in protection.

What is the pathway of B. pertussis? ›

B. pertussis is transmitted by aerosols and infects the ciliated epithelium of the airways. There is no further dissemination of the infection, but bacterial toxins produced in the respiratory tract contribute to local and systemic disease pathogenesis [10–13].

What is the main virulence factor of Bordetella pertussis? ›

Some of these factors, including pertussis toxin (PT), adenylate cyclase, tracheal cytotoxin, dermonecrotic toxin, and lipopolysaccharide, exert a toxic effect. Other virulence factors, such as filamentous hemagglutinin (FHA), fimbriae, pertactin (Prn), and PT, have been reported to mediate the adherence of B.

What is the name of the toxin produced by Bordetella pertussis that causes the loss of cilia? ›

Of the virulence factors produced by Bordetella organisms, only tracheal cytotoxin induces paralysis and destruction of respiratory ciliated epithelium, the hallmark of pertussis disease.

What is the name of the toxin produced by Bordetella pertussis that causes the loss of cilia in the lower respiratory tract? ›

These toxins include the pertussis toxin (exotoxin) that activates host cell production of cAMP to modulate cell protein synthesis regulation, a tracheal cytotoxin that causes destruction of ciliated epithelial cells, and a cell surface hemagglutinin to assist in bacterial binding to the host cells.

How do you diagnose pertussis? ›

By taking a nose and throat swab, they can test for the Bordetella pertussis bacteria in culture or a more rapid polymerase chain reaction test. Your doctor may suggest a blood test to check your white blood cell count, which, if low, signals the presence of an infection.

Which gram-negative bacilli is most common? ›

Gram-negative infections include those caused by Klebsiella, Acinetobacter, Pseudomonas aeruginosa, and E. coli., as well as many other less common bacteria.

How do you identify Bordetella pertussis? ›

Scientists consider culture the gold standard because it is the only 100% specific method for Bordetella pertussis identification. Other tests that can be performed include polymerase chain reaction (PCR) and serology.

What is the difference between pertussis IgG and IgM? ›

With blood testing, pertussis IgG antibodies will be present in those who have been vaccinated or have had a past infection. IgM and IgA antibodies may indicate recent vaccination or infection and will only be present for a short period of time (2-3 months).

Is pertussis gram positive or negative? ›

Bordetella pertussis is a small (approximately 0.8 μm by 0.4 μm), rod-shaped, coccoid, or ovoid Gram-negative bacterium that is encapsulated and does not produce spores.

Do adults need pertussis booster? ›

Any adult 19 years of age or older who has never received a dose of Tdap should get one as soon as feasible. You should administer Tdap regardless of interval since the last tetanus or diphtheria toxoid-containing vaccine. This should be followed by either a Td or Tdap shot every 10 years.

Can you get pertussis if vaccinated? ›

Can people who have been vaccinated still get whooping cough? Sometimes when vaccinated people are exposed, they get whooping cough anyway, although they usually have milder symptoms, a shorter illness, and may be less likely to spread the disease to others.

What is the safest pertussis vaccine? ›

DTaP and Tdap vaccine are safe and effective at preventing diphtheria, tetanus, and pertussis. Vaccines, like any medicine, can have side effects. The most common side effects are usually mild and go away on their own. Severe allergic reactions following vaccination are rare, but can be life threatening.

Can you get rid of pertussis without antibiotics? ›

Pertussis bacteria die off naturally after three weeks of coughing. If antibiotics are not started within that time, they are no longer recommended. Antibiotics can also be given to close contacts of persons with pertussis to prevent or lessen the symptoms.

How do you get infected with pertussis? ›

Pertussis bacteria are spread through droplets produced during coughing or sneezing. These droplets don't travel very far through the air and usually only infect persons nearby.

What is the best way to reduce pertussis? ›

The best way to prevent whooping cough is to get vaccinated. Two vaccines in the United States help prevent whooping cough: DTaP and Tdap. These vaccines also provide protection against tetanus and diphtheria.

Is pertussis toxin a hazard? ›

Pertussis toxin is an environmental hazard. Do not allow inactivated toxin to enter the drain. Solid waste including pipettes, containers, etc. that come in contact with toxin must be disinfected with 10% bleach prior to disposal in biohazard waste container.

What class of drug is pertussis? ›

Macrolides erythromycin, clarithromycin, and azithromycin* are preferred for the treatment of pertussis in persons 1 month of age and older. For persons 2 months of age and older, an alternative to macrolides is trimethoprim-sulfamethoxazole.

What class of vaccine is used for pertussis? ›

There are 2 vaccines that include protection against whooping cough: The DTaP vaccine protects young children from diphtheria, tetanus, and whooping cough. The Tdap vaccine protects preteens, teens, and adults from tetanus, diphtheria, and whooping cough.

What is pertussis more commonly known as? ›

Pertussis (Whooping Cough)

Is pertussis vaccine live or killed? ›

Killed (inactivated) vaccines are made from a protein or other small pieces taken from a virus or bacteria. The whooping cough (pertussis) vaccine is an example.

What is the difference between whooping cough and pertussis? ›

Pertussis is also called whooping cough. It is a serious lung infection caused by bacteria. It is also very contagious and causes coughing fits. Whooping cough is most serious for babies with very small airways.

What is the most common cause of pertussis? ›

Whooping cough is caused by a type of bacteria called Bordetella pertussis. It spreads from person to person. People who have pertussis usually spread it through coughing, sneezing, or breathing very close to someone. It can also sometimes be spread by touching an infected surface and then touching your nose or mouth.

What stage of pertussis is most contagious? ›

People with pertussis are most infectious during the catarrhal period and during the first two weeks after onset of the cough (approximately 21 days).

What does pertussis toxin cause? ›

Pertussis toxin (PT) is a protein-based AB5-type exotoxin produced by the bacterium Bordetella pertussis, which causes whooping cough.

What are the symptoms of pertussis toxin? ›

Early symptoms can last for 1 to 2 weeks and usually include:
  • Runny or stuffed-up nose.
  • Low-grade fever (less than 100.4°F)
  • Mild, occasional cough (babies do not do this)
  • Apnea (life-threatening pauses in breathing) and cyanosis (turning blue or purple) in babies and young children.

Does diphtheria toxin have a mechanism of action? ›

Diphtheria toxin kills cells by inhibiting eukaryotic protein synthesis, and its mechanism of action has been extensively characterized. This potent toxin inactivates elongation factor (EF-2) required for protein synthesis (Collier, 1967).

What does pertussis toxin do to cAMP? ›

Bordetella pertussis produces extracytoplasmic adenylate cyclase toxin (AC toxin) which penetrates target cells and, upon activation by host calmodulin, generates high levels of intracellular cyclic AMP (cAMP). As a result, bactericidal functions of immune effector cells are impaired.

How does the diphtheria toxin exert its action? ›

The diphtheria toxin enters and kills the cell through the combined action of its three domains: the catalytic domain (C-domain), the translocation domain (T-domain), and the receptor-binding domain (R-domain) [1].

What process and at what stage does diphtheria toxin inhibit? ›

Diphtheria toxin can be proteolytically cleaved into two fragments: an N-terminal fragment A (catalytic domain), and fragment B (transmembrane and receptor binding domains). Fragment A catalyzes the NAD+-dependent ADP-ribosylation of elongation factor 2, thereby inhibiting protein synthesis in eukaryotic cells.

What is the source of infection and mechanism of diphtheria transmission? ›

Diphtheria bacteria spread from person to person, usually through respiratory droplets, like from coughing or sneezing. People can also get sick from touching infected open sores or ulcers. Those at increased risk of getting sick include: People in the same household.

How does diphtheria toxin inhibit translation? ›

Diphtheria toxin (DT) inhibits eukaryotic translation elongation factor 2 (eEF2) by ADP-ribosylation in a fashion that requires diphthamide, a modified histidine residue on eEF2.

Does the pertussis vaccine contain the killed bacteria? ›

Standard pertussis vaccine prepared from killed whole cell Bordetella pertussis organisms has been in widespread use since the early 1950s. It has been effective in reducing the incidence of pertussis in the United States to low levels.

What are the two main way of transmission of diphtheria? ›

Diphtheria is caught by coming into contact with droplets from the respiratory tract of an infected person—particularly spread by coughing or sneezing. If the disease is cutaneous, it can be spread by coming into contact with the wounds or lesions of an infected person.

What toxins need to be neutralized in the treatment diphtheria? ›

Diphtheria is fatal in 5–10% of cases, but children under the age of five have a mortality rate of up to 20%. Treatment involves antibiotics to kill the bacteria (erythromycin or penicillin for 14 days) and administering of diphtheria antitoxin (DAT) to neutralize the effects of the toxin46.

What does diphtheria toxin mainly affect? ›

Diphtheria (dif-THEER-e-uh) is a serious bacterial infection that usually affects the mucous membranes of the nose and throat.

What immune cells are involved in pertussis? ›

pertussis infection induces innate immune cell (such as neutrophils and macrophages) activation, as well as IFN-γ-producing and IL-17-producing T cells. The Th-17 T cells are instrumental for sIgA production. The infection also results in resident memory T cells (γδ and CD4+ T cells).

Which cells are affected by pertussis? ›

Pertussis is primarily a toxin-mediated disease. The bacteria attach to the cilia of the respiratory epithelial cells, produce toxins that paralyze the cilia, and cause inflammation of the respiratory tract, which interferes with the clearing of pulmonary secretions.

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