scholarly journals DegP initiates regulated processing of filamentous hemagglutinin in Bordetella bronchiseptica

2021 ◽  
Author(s):  
Richard M Johnson ◽  
Zachary M Nash ◽  
Margaret R Dedloff ◽  
John C Shook ◽  
Peggy A Cotter
Keyword(s):  
Pathogenic Bacteria ◽  
Whooping Cough ◽  
Bordetella Bronchiseptica ◽  
Host Cells ◽  
Closely Related Species ◽  
Bacterial Surface ◽  
Filamentous Hemagglutinin ◽  
C Terminus ◽  
Cell Clearance ◽  
Culture Supernatants

Filamentous hemagglutinin (FhaB) is a critical virulence factor for both Bordetella bronchiseptica, the causal agent of whooping cough, and the closely related species Bordetella bronchiseptica. FhaB is an adhesin, suppresses inflammatory cytokine production, and protects against phagocytic cell clearance during infection. Regulated degradation of the FhaB C-terminal prodomain is required to establish a persistent infection in mice. Two proteases, CtpA in the periplasm and SphB1 on the bacterial surface, mediate FhaB processing, and we recently determined that CtpA functions before, and controls the FhaB cleavage site of, SphB1. However, the data indicate that another periplasmic protease must initiate degradation of the prodomain by removing a portion of the FhaB C terminus that inhibits CtpA-mediated degradation. Using a candidate approach, we identified DegP as the initiating protease. Deletion of degP or substitution of its predicted catalytic residue resulted in reduced creation of FHA', the main product of FhaB processing, and an accumulation of full-length FhaB in whole cell lysates. Also, FHA' was no longer released into culture supernatants in degP mutants. Alterations of the FhaB C terminus that relieve inhibition of CtpA abrogate the need for DegP, consistent with DegP functioning prior to CtpA in the processing pathway. DegP is not required for secretion of FhaB through FhaC or for adherence of the bacteria to host cells, indicating that DegP acts primarily as a protease and not a chaperone for FhaB in B. bronchiseptica. Our results highlight a role for HtrA family proteases in activation of virulence factors in pathogenic bacteria.

scholarly journals Bordetella pertussis filamentous hemagglutinin interacts with a leukocyte signal transduction complex and stimulates bacterial adherence to monocyte CR3 (CD11b/CD18).

1994 ◽  
Vol 180 (4) ◽  
pp. 1225-1233 ◽  
Author(s):  
Y Ishibashi ◽  
S Claus ◽  
D A Relman
Keyword(s):  
Signal Transduction ◽  
Bordetella Pertussis ◽  
Whooping Cough ◽  
Bacterial Pathogen ◽  
Binding Activity ◽  
Host Cells ◽  
Bacterial Surface ◽  
Filamentous Hemagglutinin ◽  
Complement Receptor 3 ◽  
Leukocyte Response

Bordetella pertussis, the causative agent of whooping cough, adheres to human monocytes/macrophages by means of a bacterial surface-associated protein, filamentous hemagglutinin (FHA) and the leukocyte integrin, complement receptor 3 (CR3, alpha M beta 2, CD11b/CD18). We show that an FHA Arg-Gly-Asp site induces enhanced B. pertussis binding to monocytes, and that this enhancement is blocked by antibodies directed against CR3. Enhancement requires a monocyte signal transduction complex, composed of leukocyte response integrin (alpha? beta 3) and integrin-associated protein (CD47). This complex is known to upregulate CR3 binding activity. Thus, a bacterial pathogen enhances its own attachment to host cells by coopting a host cell signaling pathway.

scholarly journals Filamentous Hemagglutinin of Bordetella bronchiseptica Is Required for Efficient Establishment of Tracheal Colonization

1998 ◽  
Vol 66 (12) ◽  
pp. 5921-5929 ◽  
Author(s):  
Peggy A. Cotter ◽  
Ming H. Yuk ◽  
Seema Mattoo ◽  
Brian J. Akerley ◽  
Jeff Boschwitz ◽  
...  
Keyword(s):  
Whooping Cough ◽  
Ectopic Expression ◽  
Etiologic Agent ◽  
Bordetella Bronchiseptica ◽  
Host Animal ◽  
Filamentous Hemagglutinin ◽  
Unanesthetized Animals ◽  
Necessary And Sufficient

ABSTRACT Adherence to ciliated respiratory epithelial cells is considered a critical early step in Bordetella pathogenesis. ForBordetella pertussis, the etiologic agent of whooping cough, several factors have been shown to mediate adherence to cells and cell lines in vitro. These putative adhesins include filamentous hemagglutinin (FHA), fimbriae, pertactin, and pertussis toxin. Determining the precise roles of each of these factors in vivo, however, has been difficult, due in part to the lack of natural-host animal models for use with B. pertussis. Using the closely related species Bordetella bronchiseptica, and by constructing both deletion mutation and ectopic expression mutants, we have shown that FHA is both necessary and sufficient for mediating adherence to a rat lung epithelial (L2) cell line. Using a rat model of respiratory infection, we have shown that FHA is absolutely required, but not sufficient, for tracheal colonization in healthy, unanesthetized animals. FHA was not required for initial tracheal colonization in anesthetized animals, however, suggesting that its role in establishment may be dedicated to overcoming the clearance action of the mucociliary escalator.

scholarly journals Discovery of Fibrillar Adhesins across Bacterial Species

2020 ◽  
Author(s):  
Vivian Monzon ◽  
Aleix Lafita ◽  
Alex Bateman
Keyword(s):  
Cell Surface ◽  
Bacterial Species ◽  
Domain Architecture ◽  
Surface Proteins ◽  
Host Cells ◽  
Bacterial Surface ◽  
Bacterial Phyla ◽  
C Terminus ◽  
Wide Range ◽  
Bacterial Proteomes

AbstractBackgroundFibrillar adhesins are long multidomain proteins attached at the cell surface and composed of at least one adhesive domain and multiple tandemly repeated domains, which build an elongated stalk that projects the adhesive domain beyond the bacterial cell surface. They are an important yet understudied class of proteins that mediate interactions of bacteria with their environment. This study aims to characterize fibrillar adhesins in a wide range of bacterial phyla and to identify new fibrillar adhesin-like proteins to improve our understanding of host-bacteria interactions.ResultsBy careful search for fibrillar adhesins in the literature and by computational analysis we identified 75 stalk domains and 24 adhesive domains. Based on the presence of these domains in the UniProt Reference Proteomes database, we identified and analysed 3,388 fibrillar adhesin-like proteins across species of the most common bacterial phyla. We found that the bacterial proteomes with the highest fraction of fibrillar adhesins include several known pathogens. We further enumerate the adhesive and stalk domain combinations found in nature and demonstrate that fibrillar adhesins have complex and variable domain architectures, which differ across species. By analysing the domain architecture of fibrillar adhesins we show that in Gram positive bacteria adhesive domains are mostly positioned at the N-terminus of the protein with the cell surface anchor at the C-terminus, while their positions are more variable in Gram negative bacteria. We provide an open repository of fibrillar adhesin-like proteins and domains to facilitate downstream studies of this class of bacterial surface proteins.ConclusionThis study provides a domain-based characterization of fibrillar adhesins and demonstrates that they are widely found across the main bacterial phyla. We have discovered numerous novel fibrillar adhesins and improved the understanding of how pathogens might adhere to and subsequently invade into host cells.

scholarly journals Production of Nontypeable Haemophilus influenzae HtrA by Recombinant Bordetella pertussis with the Use of Filamentous Hemagglutinin as a Carrier

2005 ◽  
Vol 73 (7) ◽  
pp. 4295-4301 ◽  
Author(s):  
Sylvie Alonso ◽  
Eve Willery ◽  
Genevieve Renauld-Mongénie ◽  
Camille Locht
Keyword(s):  
Immune Responses ◽  
Haemophilus Influenzae ◽  
Bordetella Pertussis ◽  
Whooping Cough ◽  
Etiologic Agent ◽  
Nontypeable Haemophilus Influenzae ◽  
Bacterial Surface ◽  
Filamentous Hemagglutinin ◽  
Bacterial Vector ◽  
Antibody Titers

ABSTRACT Bordetella pertussis, the etiologic agent of whooping cough, is a highly infectious human pathogen capable of inducing mucosal and systemic immune responses upon a single intranasal administration. In an attenuated, pertussis toxin (PTX)-deficient recombinant form, it may therefore constitute an efficient bacterial vector that is particularly well adapted for the delivery of heterologous antigens to the respiratory mucosa. Filamentous hemagglutinin (FHA) has been used as a carrier to present foreign antigens at the bacterial surface, thereby inducing local, systemic, and protective immune responses to these antigens in mice. Both full-length and truncated (Fha44) forms of FHA have been used for antigen presentation. To investigate the effect of the carrier (FHA or Fha44) on antibody responses to passenger antigens, we genetically fused the HtrA protein of nontypeable Haemophilus influenzae to either FHA form. The fha-htrA and Fha44 gene-htrA hybrids were expressed as single copies inserted into the chromosome of PTX-deficient B. pertussis. Both chimeras were secreted into the culture supernatants of the recombinant strains and were recognized by anti-FHA and anti-HtrA antibodies. Intranasal infection with the strain producing the FHA-HtrA hybrid led to significantly higher anti-HtrA and anti-FHA antibody titers than those obtained in mice infected with the Fha44-HtrA-producing strain. Interestingly, the B. pertussis strain producing the Fha44-HtrA chimera colonized the mouse lungs more efficiently than the parental, Fha44-producing strain and gave rise to higher anti-FHA antibody titers than those induced by the parental strain.

scholarly journals Passively Released Heme from Hemoglobin and Myoglobin Is a Potential Source of Nutrient Iron for Bordetella bronchiseptica

2007 ◽  
Vol 75 (10) ◽  
pp. 4857-4866 ◽  
Author(s):  
Jeffrey C. Mocny ◽  
John S. Olson ◽  
Terry D. Connell
Keyword(s):  
Direct Contact ◽  
Pathogenic Bacteria ◽  
Whooping Cough ◽  
Respiratory Infections ◽  
Bordetella Bronchiseptica ◽  
Common Mechanism ◽  
Atrophic Rhinitis ◽  
Dependent Manner ◽  
Potential Source ◽  
Successful Colonization

ABSTRACT Colonization by Bordetella bronchiseptica results in a variety of inflammatory respiratory infections, including canine kennel cough, porcine atrophic rhinitis, and a whooping cough-like disease in humans. For successful colonization, B. bronchiseptica must acquire iron (Fe) from the infected host. A vast amount of Fe within the host is sequestered within heme, a metalloporphyrin which is coordinately bound in hemoglobin and myoglobin. Utilization of hemoglobin and myoglobin as sources of nutrient Fe by B. bronchiseptica requires expression of BhuR, an outer membrane protein. We hypothesize that hemin is acquired by B. bronchiseptica in a BhuR-dependent manner after spontaneous loss of the metalloporphyrin from hemoglobin and/or myoglobin. Sequestration experiments demonstrated that direct contact with hemoglobin or myoglobin was not required to support growth of B. bronchiseptica in an Fe-limiting environment. Mutant myoglobins, each exhibiting a different affinity for heme, were employed to demonstrate that the rate of growth of B. bronchiseptica was directly correlated with the rate at which heme was lost from the hemoprotein. Finally, Escherichia coli cells expressing recombinant BhuR had the capacity to remove hemin from solution. Collectively, these experiments provided strong experimental support for the model that BhuR is a hemin receptor and B. bronchiseptica likely acquires heme during infection after passive loss of the metalloporphyrin from hemoglobin and/or myoglobin. These results also suggest that spontaneous hemin loss by hemoglobin and myoglobin may be a common mechanism by which many pathogenic bacteria acquire heme and heme-bound Fe.

scholarly journals Structure and Function of Hib Pili from Haemophilus influenzae Type b

2002 ◽  
Vol 184 (17) ◽  
pp. 4868-4874 ◽  
Author(s):  
Xiang-Qi Mu ◽  
Edward H. Egelman ◽  
Esther Bullitt
Keyword(s):  
Haemophilus Influenzae ◽  
Pathogenic Bacteria ◽  
Three Dimensional ◽  
Host Cells ◽  
Haemophilus Influenzae Type ◽  
Helical Axis ◽  
Type B ◽  
Environmental Niche ◽  
Bacterial Surface ◽  
And Function

ABSTRACT Pathogenic bacteria are specifically adapted to bind to their customary host. Disease is then caused by subsequent colonization and/or invasion of the local environmental niche. Initial binding of Haemophilus influenzae type b to the human nasopharynx is facilitated by Hib pili, filaments expressed on the bacterial surface. With three-dimensional reconstruction of electron micrograph images, we show that Hib pili comprise a helix 70 Å in diameter with threefold symmetry. The Hib pilus filament has 3.0 subunits per turn, with each set of three subunits translated 26.9 Å along and rotated 53 degrees about the helical axis. Amino acid sequence analysis of pilins from Hib pili and from P-pili expressed on uropathogenic Escherichia coli were used to predict the physical location of the highly variable and immunogenic region of the HifA pilin in the Hib pilus structure. Structural differences between Hib pili and P-pili suggest a difference in the strategies by which bacteria remain bound to their host cells: P-pili were shown to be capable of unwinding to five times their original length (E. Bullitt and L. Makowski, Nature 373:164-167, 1995), while damage to Hib pili occurs by slight shearing of subunits with respect to those further along the helical axis. This capacity to resist unwinding may be important for continued adherence of H. influenzae type b to the nasopharynx, where the three-stranded Hib pilus filaments provide a robust tether to withstand coughs and sneezes.

Blocking Vibrio cholerae-mediated hemagglutination with short peptide antagonists

2020 ◽  
Author(s):  
Shuaiqi Guo ◽  
Cameron Lloyd ◽  
Brett Kinrade ◽  
Mustafa Sherik ◽  
Ilja Voets ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Bacterial Infections ◽  
Pathogenic Bacteria ◽  
Host Cells ◽  
Antibiotic Resistant ◽  
X Ray Crystallography ◽  
Ice Binding ◽  
C Terminus ◽  
Erythrocyte Binding ◽  
Binding Cavity

<p>Many bacteria use repeats-in-toxin (RTX) adhesins to mediate binding to host cells and facilitate subsequent colonisation and infection by forming biofilms. Vibrio cholerae, the causative agent of cholera, uses a 230-kDa RTX adhesin, FrhA, to facilitate intestinal colonization. FrhA also mediates hemagglutination of red-blood cells (erythrocytes). Here we have demonstrated that the hemagglutination capability of FrhA is localized to a ~ 20-kDa domain near its C terminus. Bioinformatic analyses indicated this erythrocyte-binding domain (VcEBD) is 65% identical to a peptide-binding module found in the 1.5-MDa ice-binding RTX adhesin that helps its Antarctic bacterium, Marinomonas primoryensis, form symbiotic biofilms with diatoms on the underside of sea ice. This suggested that the FrhA binds V. cholerae to proteins present on the cell surface of erythrocytes. X-ray crystallography revealed that VcEBD has an oblong β-sandwich fold with a shallow, Ca<sup>2+</sup>-dependent ligand-binding cavity that can anchor a peptidyl ligand with a free terminal carboxyl group. Using a structure-guided approach, we screened a small library of ~ 60 short peptides and optimized the affinity of VcEBD’s peptidyl ligands by roughly 1,000-fold. Importantly, the high-affinity ligands are effective at blocking V. cholerae from binding to erythrocytes at nano-molar concentrations. Structures of VcEBD in complex with three different peptides further elucidated the molecular basis for their interactions, which sets the stage for the development of ligand-based antagonists that may help disrupt V. cholerae interaction with intestinal cells to prevent or treat cholera. With the spread of antibiotic-resistant pathogenic bacteria, this work sheds light on an anti-adhesion approach for combating bacterial infections without the excessive use of antibiotics.</p>

SARS-CoV-2 Biology Insights, Part IV.Antibody-Dependent Enhancement (ADE) and the tricky “Herd Immunity”: narrative review (Preprint)

2020 ◽  
Author(s):  
Laura Lafon-Hughes
Keyword(s):  
Viral Infection ◽  
Viral Entry ◽  
Neutralizing Antibodies ◽  
Vaccine Development ◽  
Whooping Cough ◽  
Common Knowledge ◽  
Herd Immunity ◽  
Life Quality ◽  
Host Cells ◽  
System P

BACKGROUND It is common knowledge that vaccination has improved our life quality and expectancy since it succeeded in achieving almost eradication of several diseases including chickenpox (varicella), diphtheria, hepatitis A and B, measles, meningococcal, mumps, pneumococcal, polio, rotavirus, rubella, tetanus and whooping cough (pertussis) Vaccination success is based on vaccine induction of neutralizing antibodies that help fight the infection (e.g. by a virus), preventing the disease. Conversely, Antibody-dependent enhancement (ADE) of a viral infection occurs when anti-viral antibodies facilitate viral entry into host cells and enhance viral infection in these cells. ADE has been previously studied in Dengue and HIV viruses and explains why a second infection with Dengue can be lethal. As already reviewed in Part I and Part II, SARS-Cov-2 shares with HIV not only 4 sequences in the Spike protein but also the capacity to attack the immune system. OBJECTIVE As HIV presents ADE, we wondered whether this was also the case regarding SARS-CoV-2. METHODS A literature review was done through Google. RESULTS SARS-CoV-2 presents ADE. As SARS, which does not have the 4 HIV-like inserts, has the same property, ADE would not be driven by the HIV-like spike sequences. CONCLUSIONS ADE can explain the failure of herd immunity-based strategies and will also probably hamper anti-SARS-CoV-2 vaccine development. As reviewed in Part I, there fortunately are promising therapeutic strategies for COVID-19, which should be further developed. In the meantime, complementary countermeasures to protect mainly the youth from this infection are presented to be discussed in Part V Viewpoint.

Airborne-induced experimental Bordetella bronchiseptica pneumonia in Strain 13 guineapigs

1987 ◽  
Vol 21 (3) ◽  
pp. 226-232 ◽  
Author(s):  
C. J. Trahan ◽  
E. H. Stephenson ◽  
J. W. Ezzell ◽  
W. C. Mitchell
Keyword(s):  
Weight Loss ◽  
Lethal Dose ◽  
Clinical Signs ◽  
Model System ◽  
Bordetella Bronchiseptica ◽  
Host Cells ◽  
Nasal Discharge ◽  
Disease Pathogenesis ◽  
Infectious Dose ◽  
Colony Forming Units

To evaluate the efficacy of a commercial bacterial vaccine in protecting Strain 13 guineapigs against fatal Bordetella bronchiseptica pneumonia, it was necessary to establish the infectivity and disease pathogenesis induced by virulent organisms. When guineapigs were exposed to small-particle aerosols of varying concentrations of virulent B. bronchiseptica, a spectrum of disease was produced that ranged from inapparent illness to fulminant bronchopneumonia. Clinical signs began by day 4 after exposure, and were evidenced by anorexia, weight loss, respiratory distress and serous to purulent nasal discharge. Pathological alterations were limited to the respiratory system. Moribund animals exhibited a suppurative necrotizing bronchopneumonia and necrotizing tracheitis. In animals that survived the challenge, the bacteria were eliminated from the lungs by day 28 but continued to persist in the laryngeal area and the trachea. The median infectious dose and the median lethal dose were estimated to be 4 colony-forming units (CFU) and 1314 CFU respectively. These data suggest that the guineapig will be a valuable model system in which to study interactions between Bordetella species and host cells as well as to evaluate potential B. bronchiseptica immunogens.

scholarly journals Virtual Screening for Potential Inhibitors of Human Hexokinase II for the Development of Anti-Dengue Therapeutics

BioTech ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 1
Author(s):  
Suriyea Tanbin ◽  
Fazia Adyani Ahmad Fuad ◽  
Azzmer Azzar Abdul Hamid
Keyword(s):  
Hydrogen Bonds ◽  
Binding Energies ◽  
Host Cells ◽  
Dynamics Simulation ◽  
Hexokinase Ii ◽  
Tropical Regions ◽  
Lipinski’S Rule ◽  
Viral Propagation ◽  
C Terminus ◽  
Similarity Scores

Dengue fever, which is a disease caused by the dengue virus (DENV), is a major unsolved issue in many tropical and sub-tropical regions of the world. The absence of treatment that effectively prevent further viral propagation inside the human’s body resulted in a high number of deaths globally each year. Thus, novel anti-dengue therapies are required for effective treatment. Human hexokinase II (HKII), which is the first enzyme in the glycolytic pathway, is an important drug target due to its significant impact on viral replication and survival in host cells. In this study, 23.1 million compounds were computationally-screened against HKII using the Ultrafast Shape Recognition with a CREDO Atom Types (USRCAT) algorithm. In total, 300 compounds with the highest similarity scores relative to three reference molecules, known as Alpha-D-glucose (GLC), Beta-D-glucose-6-phosphate (BG6), and 2-deoxyglucose (2DG), were aligned. Of these 300 compounds, 165 were chosen for further structure-based screening, based on their similarity scores, ADME analysis, the Lipinski’s Rule of Five, and virtual toxicity test results. The selected analogues were subsequently docked against each domain of the HKII structure (PDB ID: 2NZT) using AutoDock Vina programme. The three top-ranked compounds for each query were then selected from the docking results based on their binding energy, the number of hydrogen bonds formed, and the specific catalytic residues. The best docking results for each analogue were observed for the C-terminus of Chain B. The top-ranked analogues of GLC, compound 10, compound 26, and compound 58, showed predicted binding energies of −7.2, −7.0, and −6.10 kcal/mol and 7, 5, and 2 hydrogen bonds, respectively. The analogues of BG6, compound 30, compound 36, and compound 38, showed predicted binding energies of −7.8, −7.4, and −7.0 kcal/mol and 11, 9, and 5 hydrogen bonds, while the top three analogues of 2DG, known as compound 1, compound 4, and compound 31, showed predicted binding energies of −6.8, −6.3, and −6.3 kcal/mol and 4, 3, and 1 hydrogen bonds, sequentially. The highest-ranked compounds in the docking analysis were then selected for molecular dynamics simulation, where compound 10, compound 30, and compound 1, which are the analogues of GLC, BG6, and 2DG, have shown strong protein-ligand stability with an RMSD value of ±5.0 A° with a 5 H bond, ±4.0 A° with an 8 H bond, and ±0.5 A° with a 2 H bond, respectively, compared to the reference molecules throughout the 20 ns simulation time. Therefore, by using the computational studies, we proposed novel compounds, which may act as potential drugs against DENV by inhibiting HKII’s activity.

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