8 results on '"Wouter S. P. Jong"'
Search Results
2. Overexpression of the Bam Complex Improves the Production of Chlamydia trachomatis MOMP in the E. coli Outer Membrane
- Author
-
Dung T. Huynh, Wouter S. P. Jong, Gregory M. Koningstein, Peter van Ulsen, Joen Luirink, Molecular Microbiology, AIMMS, LaserLaB - Analytical Chemistry and Spectroscopy, and LaserLaB - Molecular Biophysics
- Subjects
Organic Chemistry ,E. coli ,Chlamydia trachomatis major outer membrane protein ,OMV-based vaccine ,General Medicine ,Catalysis ,Computer Science Applications ,Inorganic Chemistry ,SDG 3 - Good Health and Well-being ,Outer membrane protein ,β-barrel assembly machinery ,outer membrane protein ,Physical and Theoretical Chemistry ,Molecular Biology ,Spectroscopy - Abstract
A licensed Chlamydia trachomatis (Ct) vaccine is not yet available. Recombinant Chlamydia trachomatis major outer membrane protein (Ct-MOMP), the most abundant constituent of the chlamydial outer membrane complex, is considered the most attractive candidate for subunit-based vaccine formulations. Unfortunately, Ct-MOMP is difficult to express in its native structure in the E. coli outer membrane (OM). Here, by co-expression of the Bam complex, we improved the expression and localization of recombinant Ct-MOMP in the E. coli OM. Under these conditions, recombinant Ct-MOMP appeared to assemble into a β-barrel conformation and express domains at the cell surface indicative of correct folding. The data indicate that limited availability of the Bam complex can be a bottleneck for the production of heterologous OM vaccine antigens, information that is also relevant for strategies aimed at producing recombinant OMV-based vaccines.
- Published
- 2022
3. Exploring metal availability in the natural niche of Streptococcus pneumoniae to discover potential vaccine antigens
- Author
-
H. Bart van den Berg van Saparoea, Daniela M. Ferreira, Thomas H. A. Ederveen, Irma Joosten, Joen Luirink, Lucille F. van Beek, Diane Houben, Kristin Surmann, Elena Mitsi, Uwe Völker, Frank Schmidt, Christian Hentschker, Dimitri A. Diavatopoulos, Fred van Opzeeland, Christa E. van der Gaast-de Jongh, Wouter S. P. Jong, Marien I. de Jonge, Molecular Microbiology, AIMMS, and LaserLaB - Molecular Biophysics
- Subjects
Serotype ,Male ,wc_100 ,protein antigens ,lnfectious Diseases and Global Health Radboud Institute for Molecular Life Sciences [Radboudumc 4] ,Infectious and parasitic diseases ,RC109-216 ,medicine.disease_cause ,Pneumococcal Vaccines ,Mice ,Nasopharynx ,Colonization ,0303 health sciences ,Transmission (medicine) ,nasal fluid ,wc_217 ,Middle Aged ,Antibodies, Bacterial ,Infectious Diseases ,Streptococcus pneumoniae ,Metals ,Proteome ,Female ,Pneumonia (non-human) ,Inflammatory diseases Radboud Institute for Molecular Life Sciences [Radboudumc 5] ,Research Article ,Research Paper ,Microbiology (medical) ,Adult ,Immunology ,Biology ,Microbiology ,transition metals ,Pneumococcal Infections ,qw_805 ,03 medical and health sciences ,Young Adult ,All institutes and research themes of the Radboud University Medical Center ,SDG 3 - Good Health and Well-being ,medicine ,Animals ,Humans ,Molecular Biology ,030304 developmental biology ,Antigens, Bacterial ,Proteomic Profile ,030306 microbiology ,in vivo-mimicking ,Membrane Proteins ,medicine.disease ,colonization ,Nasal Lavage Fluid ,Culture Media ,Mice, Inbred C57BL ,Disease Models, Animal ,Parasitology ,qw_142 ,Oligopeptide binding - Abstract
Nasopharyngeal colonization by Streptococcus pneumoniae is a prerequisite for pneumococcal transmission and disease. Current vaccines protect only against disease and colonization caused by a limited number of serotypes, consequently allowing serotype replacement and transmission. Therefore, the development of a broadly protective vaccine against colonization, transmission and disease is desired but requires a better understanding of pneumococcal adaptation to its natural niche. Hence, we measured the levels of free and protein-bound transition metals in human nasal fluid, to determine the effect of metal concentrations on the growth and proteome of S. pneumoniae. Pneumococci cultured in medium containing metal levels comparable to nasal fluid showed a highly distinct proteomic profile compared to standard culture conditions, including the increased abundance of nine conserved, putative surface-exposed proteins. AliA, an oligopeptide binding protein, was identified as the strongest protective antigen, demonstrated by the significantly reduced bacterial load in a murine colonization and a lethal mouse pneumonia model, highlighting its potential as vaccine antigen.
- Published
- 2020
4. Highly conserved nucleotide phosphatase essential for membrane lipid homeostasis inStreptococcus pneumoniae
- Author
-
Kirsten Kuipers, Marketa Martinkova, Hanka Venselaar, Aldert Zomer, Václav Martínek, Wouter S. P. Jong, Manfred Rohde, Jan-Willem Veening, Samantha L. van der Beek, Hester J. Bootsma, Marien I. de Jonge, and Clement Gallay
- Subjects
0301 basic medicine ,chemistry.chemical_classification ,biology ,ATP synthase ,030106 microbiology ,Phosphatase ,Mutant ,Virulence ,Bacillus subtilis ,biology.organism_classification ,medicine.disease_cause ,Microbiology ,03 medical and health sciences ,Enzyme ,Biochemistry ,chemistry ,Lipid biosynthesis ,medicine ,biology.protein ,Molecular Biology ,Escherichia coli - Abstract
Proteins belonging to the DHH family, a member of the phosphoesterase superfamily, are produced by most bacterial species. While some of these proteins are well studied in Bacillus subtilis and Escherichia coli, their functions in Streptococcus pneumoniae remain unclear. Recently, the highly conserved DHH subfamily 1 protein PapP (SP1298) has been reported to play an important role in virulence. Here, we provide a plausible explanation for the attenuated virulence of the papP mutant. Recombinant PapP specifically hydrolyzed nucleotides 3'-phosphoadenosine-5'-phosphate (pAp) and 5'-phosphoadenylyl-(3'->5')-adenosine (pApA). Deletion of papP, potentially leading to pAp/pApA accumulation, resulted in morphological defects and mis-localization of several cell division proteins. Incubation with both polar solvent and detergent led to robust killing of the papP mutant, indicating that membrane integrity is strongly affected. This is in line with previous studies showing that pAp inhibits the ACP synthase, an essential enzyme involved in lipid precursor production. Remarkably, partial inactivation of the lipid biosynthesis pathway, by inhibition of FabF or depletion of FabH, phenocopied the papP mutant. We conclude that pAp and pApA phosphatase activity of PapP is required for maintenance of membrane lipid homeostasis providing an explanation how inactivation of this protein may attenuate pneumococcal virulence.
- Published
- 2016
5. Comparing autotransporter β-domain configurations for their capacity to secrete heterologous proteins to the cell surface
- Author
-
Maaike Schillemans, Peter van Ulsen, Wouter S. P. Jong, Joen Luirink, Corinne M. ten Hagen-Jongman, Graduate School, ACS - Microcirculation, Molecular Microbiology, AIMMS, LaserLaB - Molecular Biophysics, and LaserLaB - Analytical Chemistry and Spectroscopy
- Subjects
0301 basic medicine ,Physiology ,Cell Membranes ,lcsh:Medicine ,Biochemistry ,Membrane Fusion ,law.invention ,Cell membrane ,Cell Fusion ,law ,Medicine and Health Sciences ,Urea ,lcsh:Science ,Multidisciplinary ,biology ,Membrane transport protein ,Chemistry ,Organic Compounds ,Escherichia coli Proteins ,Proteases ,Recombinant Proteins ,Cell biology ,Enzymes ,Body Fluids ,medicine.anatomical_structure ,Blood ,Physical Sciences ,Recombinant DNA ,Cellular Structures and Organelles ,Anatomy ,Bacterial outer membrane ,Research Article ,Cell Physiology ,030106 microbiology ,Protein domain ,03 medical and health sciences ,medicine ,Escherichia coli ,Secretion ,lcsh:R ,Cell Membrane ,Organic Chemistry ,Chemical Compounds ,Membrane Transport Proteins ,Biology and Life Sciences ,Membrane Proteins ,Proteins ,Cell Biology ,Blood Serum ,Outer Membrane Proteins ,Protein tertiary structure ,030104 developmental biology ,biology.protein ,Enzymology ,lcsh:Q ,Physiological Processes ,Immune Serum ,Autotransporters - Abstract
Monomeric autotransporters have been extensively used for export of recombinant proteins to the cell surface of Gram-negative bacteria. A bottleneck in the biosynthesis of such constructs is the passage of the outer membrane, which is facilitated by the β-domain at the C terminus of an autotransporter in conjunction with the Bam complex in the outer membrane. We have evaluated eight β-domain constructs for their capacity to secrete fused proteins to the cell surface. These constructs derive from the monomeric autotransporters Hbp, IgA protease, Ag43 and EstA and the trimeric autotransporter Hia, which all were selected because they have been previously used for secretion of recombinant proteins. We fused three different protein domains to the eight β-domain constructs, being a Myc-tag, the Hbp passenger and a nanobody or VHH domain, and assessed expression, membrane insertion and surface exposure. Our results show that expression levels differed considerably between the constructs tested. The constructs that included the β-domains of Hbp and IgA protease appeared the most efficient and resulted in expression levels that were detectable on Coomassie-stained SDS-PAGE gels. The VHH domain appeared the most difficult fusion partner to export, probably due to its complex immunoglobulin-like structure with a tertiary structure stabilized by an intramolecular disulfide bond. Overall, the Hbp β-domain compared favorably in exporting the fused recombinant proteins, because it showed in every instance tested a good level of expression, stable membrane insertion and clear surface exposure.
- Published
- 2017
6. Highly conserved nucleotide phosphatase essential for membrane lipid homeostasis in Streptococcus pneumoniae
- Author
-
Kirsten, Kuipers, Clement, Gallay, Václav, Martínek, Manfred, Rohde, Markéta, Martínková, Samantha L, van der Beek, Wouter S P, Jong, Hanka, Venselaar, Aldert, Zomer, Hester, Bootsma, Jan-Willem, Veening, and Marien I, de Jonge
- Subjects
Binding Sites ,Virulence ,Adenine Nucleotides ,Nucleotides ,Phosphoric Monoester Hydrolases ,DEAD-box RNA Helicases ,Membrane Lipids ,Structure-Activity Relationship ,Streptococcus pneumoniae ,Bacterial Proteins ,Mutation ,Homeostasis ,Protein Binding ,Sequence Deletion - Abstract
Proteins belonging to the DHH family, a member of the phosphoesterase superfamily, are produced by most bacterial species. While some of these proteins are well studied in Bacillus subtilis and Escherichia coli, their functions in Streptococcus pneumoniae remain unclear. Recently, the highly conserved DHH subfamily 1 protein PapP (SP1298) has been reported to play an important role in virulence. Here, we provide a plausible explanation for the attenuated virulence of the papP mutant. Recombinant PapP specifically hydrolyzed nucleotides 3'-phosphoadenosine-5'-phosphate (pAp) and 5'-phosphoadenylyl-(3'-5')-adenosine (pApA). Deletion of papP, potentially leading to pAp/pApA accumulation, resulted in morphological defects and mis-localization of several cell division proteins. Incubation with both polar solvent and detergent led to robust killing of the papP mutant, indicating that membrane integrity is strongly affected. This is in line with previous studies showing that pAp inhibits the ACP synthase, an essential enzyme involved in lipid precursor production. Remarkably, partial inactivation of the lipid biosynthesis pathway, by inhibition of FabF or depletion of FabH, phenocopied the papP mutant. We conclude that pAp and pApA phosphatase activity of PapP is required for maintenance of membrane lipid homeostasis providing an explanation how inactivation of this protein may attenuate pneumococcal virulence.
- Published
- 2015
7. An autotransporter display platform for the development of multivalent recombinant bacterial vector vaccines
- Author
-
Wouter S. P. Jong, David Vikström, Carolien E. van de Sandt, Frank Follmann, Peter Andersen, Else Marie Agger, Joen Luirink, Jan-Willem de Gier, Nicole N. van der Wel, Guus F. Rimmelzwaan, Corinne M. ten Hagen-Jongman, Karin de Punder, Maria H. Daleke-Schermerhorn, Virology, Molecular Microbiology, AIMMS, and LaserLaB - Analytical Chemistry and Spectroscopy
- Subjects
Salmonella typhimurium ,Antigen delivery ,Heterologous ,Chlamydia trachomatis ,Bioengineering ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,law.invention ,Microbiology ,Bacterial genetics ,Multivalent ,03 medical and health sciences ,SDG 3 - Good Health and Well-being ,law ,Endopeptidases ,Escherichia coli ,Influenza A virus ,medicine ,Secretion ,Recombinant live vaccine ,Antigens, Viral ,Bacterial Secretion Systems ,030304 developmental biology ,Antigens, Bacterial ,0303 health sciences ,biology ,030306 microbiology ,Research ,Mycobacterium tuberculosis ,Surface display ,biology.organism_classification ,3. Good health ,Bacterial vaccine ,Influenza Vaccines ,Autotransporter ,Salmonella enterica ,Bacterial Vaccines ,Recombinant DNA ,Biotechnology - Abstract
Background The Autotransporter pathway, ubiquitous in Gram-negative bacteria, allows the efficient secretion of large passenger proteins via a relatively simple mechanism. Capitalizing on its crystal structure, we have engineered the Escherichia coli autotransporter Hemoglobin protease (Hbp) into a versatile platform for secretion and surface display of multiple heterologous proteins in one carrier molecule. Results As proof-of-concept, we demonstrate efficient secretion and high-density display of the sizeable Mycobacterium tuberculosis antigens ESAT6, Ag85B and Rv2660c in E. coli simultaneously. Furthermore, we show stable multivalent display of these antigens in an attenuated Salmonella Typhimurium strain upon chromosomal integration. To emphasize the versatility of the Hbp platform, we also demonstrate efficient expression of multiple sizeable antigenic fragments from Chlamydia trachomatis and the influenza A virus at the Salmonella cell surface. Conclusions The successful efficient cell surface display of multiple antigens from various pathogenic organisms highlights the potential of Hbp as a universal platform for the development of multivalent recombinant bacterial vector vaccines. Electronic supplementary material The online version of this article (doi:10.1186/s12934-014-0162-8) contains supplementary material, which is available to authorized users.
- Published
- 2014
8. Extracellular production of recombinant proteins using bacterial autotransporters
- Author
-
Joen Luirink, Ana Saurí, Wouter S. P. Jong, Molecular Microbiology, AIMMS, and LaserLaB - Analytical Chemistry and Spectroscopy
- Subjects
Virulence Factors ,Biomedical Engineering ,Heterologous ,Virulence ,Bioengineering ,Biology ,medicine.disease_cause ,Models, Biological ,Microbiology ,law.invention ,Bacterial Proteins ,law ,Extracellular ,medicine ,Escherichia coli ,Secretion ,Cell Membrane ,Recombinant Proteins ,Cell biology ,Periplasm ,Recombinant DNA ,Cell envelope ,Extracellular Space ,Biotechnology ,Autotransporters - Abstract
Escherichia coli is still a very popular host for the production of recombinant proteins at an analytical or industrial scale. Secretion of the proteins into the culture medium or display at the cell surface would be preferred in many applications but is hampered by the complex two-layered cell envelope. The autotransporter pathway is used by E. coli to secrete virulence factors via a relatively simple but efficient and specific mechanism. Here we discuss recent progress in the structural and mechanistic analysis of this pathway and the implications for future development of a versatile platform for secretion and display of heterologous proteins. © 2010 Elsevier Ltd.
- Published
- 2010
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.