inameus

• Meuskens, Ina; Leo, Jack Christopher & Linke, Dirk (2024). Recombinant Expression and Overproduction of Transmembrane β-Barrel Proteins. I Raffaele, Ieva (Red.), Transmembrane β-Barrel Proteins. Humana Press. ISSN 9781071637333. s. 31–41. doi: 10.1007/978-1-0716-3734-0_2. Fulltekst i vitenarkiv
• Meuskens, Ina; Kristiansen, Per Eugen; Bardiaux, Benjamin; Koynarev, Vladimir Rosenov; Hatlem, Daniel & Prydz, Kristian [Vis alle 9 forfattere av denne artikkelen] (2023). A poly-proline II helix in YadA from Yersinia enterocolitica serotype O:9 facilitates heparin binding through electrostatic interactions. The FEBS Journal. ISSN 1742-464X. 291(4), s. 761–777. doi: 10.1111/febs.17001. Fulltekst i vitenarkiv Vis sammendrag

  Poly‐proline II helices are secondary structure motifs frequently found in ligand‐binding sites. They exhibit increased flexibility and solvent exposure compared to the strongly hydrogen‐bonded α‐helices or β‐strands and can therefore easily be misinterpreted as completely unstructured regions with an extremely high rotational freedom. Here, we show that the adhesin YadA of Yersinia enterocolitica serotype O:9 contains a poly‐proline II helix interaction motif in the N‐terminal region. The motif is involved in the interaction of YadA O:9 with heparin, a host glycosaminoglycan. We show that the basic residues within the N‐terminal motif of YadA are required for electrostatic interactions with the sulfate groups of heparin. Biophysical methods including CD spectroscopy, solution‐state NMR and SAXS all independently support the presence of a poly‐proline helix allowing YadA O:9 binding to the rigid heparin. Lastly, we show that host cells deficient in sulfation of heparin and heparan sulfate are not targeted by YadA O:9 ‐mediated adhesion. We speculate that the YadA O:9 –heparin interaction plays an important and highly strain‐specific role in the pathogenicity of Yersinia enterocolitica serotype O:9.

• Meuskens, Ina; Leva-Bueno, Juan; Millner, Paul; Schütz, Monika; Peyman, Sally & Linke, Dirk (2022). The Trimeric Autotransporter Adhesin YadA of Yersinia enterocolitica Serotype O:9 Binds Glycan Moieties. Frontiers in Microbiology. 12. doi: 10.3389/fmicb.2021.738818. Fulltekst i vitenarkiv Vis sammendrag

  Yersinia adhesin A (YadA) is a key virulence factor of Yersinia enterocolitica and Yersinia pseudotuberculosis . YadA is a trimeric autotransporter adhesin, a class of adhesins that have been shown to enable many Gram-negative pathogens to adhere to/interact with the host extracellular matrix proteins such as collagen, vitronectin, and fibronectin. Here, we show for the first time that YadA of Yersinia enterocolitica serotype O:9 not only interacts with proteinaceous surface molecules but can also attach directly to glycan moieties. We show that YadA from Y. enterocolitica serotype O:9 does not interact with the vitronectin protein itself but exclusively with its N-linked glycans. We also show that YadA can target other glycan moieties as found in heparin, for example. So far, little is known about specific interactions between bacterial autotransporter adhesins and glycans. This could potentially lead to new antimicrobial treatment strategies, as well as diagnostic applications.

• Leva-Bueno, Juan; Meuskens, Ina; Linke, Dirk; Millner, Paul & Peyman, Sally A. (2022). A novel, proof-of-concept electrochemical impedimetric biosensor based on extracellular matrix protein-adhesin interaction. Sensors & Diagnostics. 1(5), s. 1003–1013. doi: 10.1039/d2sd00075j. Fulltekst i vitenarkiv Vis sammendrag

  This work presents a novel perspective for electrochemical biosensors based on the detection of the interaction between extracellular matrix proteins and bacterial adhesins. Two types of impedimetric biosensors using collagen as a bioreceptor were investigated for their ability to detect pathogen adhesion using a well-characterised system of collagen-binding to recombinant E. coli expressing adhesin. First, a collagen–polymer-matrix-based biosensor was shown to detect whole bacteria in buffer media over the range 8 × 105 cfu to 8 × 107 cfu in a 10 μL sample. Second, a step-by-step full optimised biosensor based on direct collagen attachment showed bacterial detection over the range of 8 × 104 cfu to 8 × 107 cfu in a 10 μL sample. Electrochemical impedance spectroscopy and cyclic voltammetry were employed to assess the binding, which was corroborated by fluorescence binding assays. Up to now, a wide range of bioreceptors have been used in biosensor fabrication, including antibodies, oligonucleotides, phages and molecularly imprinted polymers. In this study, we show that extracellular matrix proteins can also be used for whole pathogen recognition by binding to adhesins.

• Belkum, Alex van; Almeida, Carina; Bardiaux, Benjamin; Barrass, Sarah V.; Butcher, Sarah J. & Çaykara, Tuğçe [Vis alle 34 forfattere av denne artikkelen] (2021). Host-Pathogen Adhesion as the Basis of Innovative Diagnostics for Emerging Pathogens. Diagnostics (Basel). 11(7), s. 1–22. doi: 10.3390/diagnostics11071259. Fulltekst i vitenarkiv Vis sammendrag

  Infectious diseases are an existential health threat, potentiated by emerging and re-emerging viruses and increasing bacterial antibiotic resistance. Targeted treatment of infectious diseases requires precision diagnostics, especially in cases where broad-range therapeutics such as antibiotics fail. There is thus an increasing need for new approaches to develop sensitive and specific in vitro diagnostic (IVD) tests. Basic science and translational research are needed to identify key microbial molecules as diagnostic targets, to identify relevant host counterparts, and to use this knowledge in developing or improving IVD. In this regard, an overlooked feature is the capacity of pathogens to adhere specifically to host cells and tissues. The molecular entities relevant for pathogen–surface interaction are the so-called adhesins. Adhesins vary from protein compounds to (poly-)saccharides or lipid structures that interact with eukaryotic host cell matrix molecules and receptors. Such interactions co-define the specificity and sensitivity of a diagnostic test. Currently, adhesin-receptor binding is typically used in the pre-analytical phase of IVD tests, focusing on pathogen enrichment. Further exploration of adhesin–ligand interaction, supported by present high-throughput “omics” technologies, might stimulate a new generation of broadly applicable pathogen detection and characterization tools. This review describes recent results of novel structure-defining technologies allowing for detailed molecular analysis of adhesins, their receptors and complexes. Since the host ligands evolve slowly, the corresponding adhesin interaction is under selective pressure to maintain a constant receptor binding domain. IVD should exploit such conserved binding sites and, in particular, use the human ligand to enrich the pathogen. We provide an inventory of methods based on adhesion factors and pathogen attachment mechanisms, which can also be of relevance to currently emerging pathogens, including SARS-CoV-2, the causative agent of COVID-19.

• Wimmi, Stephan; Balinovic, Alexander; Jeckel, Hannah; Selinger, Lisa; Lampaki, Dimitrios & Eisemann, Emma [Vis alle 11 forfattere av denne artikkelen] (2021). Dynamic relocalization of cytosolic type III secretion system components prevents premature protein secretion at low external pH. Nature Communications. 12(1), s. 1–14. doi: 10.1038/s41467-021-21863-4. Fulltekst i vitenarkiv Vis sammendrag

  Abstract Many bacterial pathogens use a type III secretion system (T3SS) to manipulate host cells. Protein secretion by the T3SS injectisome is activated upon contact to any host cell, and it has been unclear how premature secretion is prevented during infection. Here we report that in the gastrointestinal pathogens Yersinia enterocolitica and Shigella flexneri , cytosolic injectisome components are temporarily released from the proximal interface of the injectisome at low external pH, preventing protein secretion in acidic environments, such as the stomach. We show that in Yersinia enterocolitica , low external pH is detected in the periplasm and leads to a partial dissociation of the inner membrane injectisome component SctD, which in turn causes the dissociation of the cytosolic T3SS components. This effect is reversed upon restoration of neutral pH, allowing a fast activation of the T3SS at the native target regions within the host. These findings indicate that the cytosolic components form an adaptive regulatory interface, which regulates T3SS activity in response to environmental conditions.

• Meuskens, Ina; Saragliadis, Athanasios; Leo, Jack Christopher & Linke, Dirk (2019). Type V secretion systems: An overview of passenger domain functions. Frontiers in Microbiology. 10, s. 1–19. doi: 10.3389/fmicb.2019.01163. Fulltekst i vitenarkiv Vis sammendrag

  Bacteria secrete proteins for different purposes such as communication, virulence functions, adhesion to surfaces, nutrient acquisition, or growth inhibition of competing bacteria. For secretion of proteins, Gram-negative bacteria have evolved different secretion systems, classified as secretion systems I through IX to date. While some of these systems consist of multiple proteins building a complex spanning the cell envelope, the type V secretion system, the subject of this review, is rather minimal. Proteins of the Type V secretion system are often called autotransporters (ATs). In the simplest case, a type V secretion system consists of only one polypeptide chain with a β-barrel translocator domain in the membrane, and an extracellular passenger or effector region. Depending on the exact domain architecture of the protein, type V secretion systems can be further separated into sub-groups termed type Va through e, and possibly another recently identified subtype termed Vf. While this classification works well when it comes to the architecture of the proteins, this is not the case for the function(s) of the secreted passenger. In this review, we will give an overview of the functions of the passengers of the different AT classes, shedding more light on the variety of functions carried out by type V secretion systems.

• Meuskens, Ina; Michalik, Marcin; Chauhan, Nandini; Linke, Dirk & Leo, Jack Christopher (2017). A New Strain Collection for Improved Expression of Outer Membrane Proteins. Frontiers in Cellular and Infection Microbiology. 7. doi: 10.3389/fcimb.2017.00464. Fulltekst i vitenarkiv Vis sammendrag

  Almost all integral membrane proteins found in the outer membranes of Gram-negative bacteria belong to the transmembrane β-barrel family. These proteins are not only important for nutrient uptake and homeostasis, but are also involved in such processes as adhesion, protein secretion, biofilm formation, and virulence. As surface exposed molecules, outer membrane β-barrel proteins are also potential drug and vaccine targets. High production levels of heterologously expressed proteins are desirable for biochemical and especially structural studies, but over-expression and subsequent purification of membrane proteins, including outer membrane proteins, can be challenging. Here, we present a set of deletion mutants derived from E. coli BL21(DE3) designed for the over-expression of recombinant outer membrane proteins. These strains harbor deletions of four genes encoding abundant β-barrel proteins in the outer membrane (OmpA, OmpC, OmpF, and LamB), both single and in all combinations of double, triple, and quadruple knock-outs. The sequences encoding these outer membrane proteins were deleted completely, leaving only a minimal scar sequence, thus preventing the possibility of genetic reversion. Expression tests in the quadruple mutant strain with four test proteins, including a small outer membrane β-barrel protein and variants thereof as well as two virulence-related autotransporters, showed significantly improved expression and better quality of the produced proteins over the parent strain. Differences in growth behavior and aggregation in the presence of high salt were observed, but these phenomena did not negatively influence the expression in the quadruple mutant strain when handled as we recommend. The strains produced in this study can be used for outer membrane protein production and purification, but are also uniquely useful for labeling experiments for biophysical measurements in the native membrane environment.

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• Meuskens, Ina; Kristiansen, Per Eugen; Bardiaux, Benjamin; Koynarev, Vladimir Rosenov; Hatlem, Daniel & Prydz, Kristian [Vis alle 9 forfattere av denne artikkelen] (2023). Strain variations in a bacterial adhesin lead to different binding partners in the pathogen Yersinia enterocolitica. Fulltekst i vitenarkiv
• Leva, Juan; Meuskens, Ina; Linke, Dirk; Peyman, Sally & Millner, Paul (2021). Novel Electrochemical Biosensor for Pathogen Detection Based on Extracellular Matrix (ECM) Protein - Adhesin Interaction. Fulltekst i vitenarkiv
• Meuskens, Ina & Linke, Dirk (2021). The Trimeric Autotransporter Adhesin YadA of Yersinia Binds to Glycan Moieties. Fulltekst i vitenarkiv
• Wimmi, Stephan; Balinovic, Alexander; Jeckel, Hannah; Selinger, Lisa; Lampaki, Dimitrios & Eisemann, Emma [Vis alle 11 forfattere av denne artikkelen] (2021). Author Correction: Dynamic relocalization of cytosolic type III secretion system components prevents premature protein secretion at low external pH (Nature Communications, (2021), 12, 1, (1625), 10.1038/s41467-021-21863-4). Nature Communications. 12(1). doi: 10.1038/s41467-021-23080-5. Fulltekst i vitenarkiv
• Meuskens, Ina; Leva-Bueno, Juan; Millner, Paul; Schütz, Monika; Peyman, Sally & Linke, Dirk (2021). The Trimeric Autotransporter Adhesin YadA of Yersinia binds to glycan moieties. Fulltekst i vitenarkiv
• Leva-Bueno, Juan; Meuskens, Ina; Linke, Dirk; Millner, Paul & Peyman, Sally (2021). Impedimetric biosensor based on extracellular matrix protein-adhesin interaction. Fulltekst i vitenarkiv
• Meuskens, Ina; Michalik, Marcin; Chauhan, Nandini; Linke, Dirk & Leo, Jack Christopher (2020). Corrigendum: A New Strain Collection for Improved Expression of Outer Membrane Proteins (Frontiers in Cellular and Infection Microbiology, (2017), 7, (464), 10.3389/fcimb.2017.00464). Frontiers in Cellular and Infection Microbiology. 10. doi: 10.3389/fcimb.2020.00220. Fulltekst i vitenarkiv
• Meuskens, Ina; Higson, Seamus & Linke, Dirk (2019). Innovative training networks: a new way of collaboration-propped PhD training. Medical Microbiology and Immmunology. ISSN 0300-8584. 209, s. 215–216. doi: 10.1007/s00430-019-00647-0. Fulltekst i vitenarkiv
• Leva, Juan; Meuskens, Ina; Gibson, Timothy M.; Linke, Dirk & Millner, Paul (2019). Biosensors for pathogen detection. Fulltekst i vitenarkiv
• Meuskens, Ina; Leva, Juan; Millner, Paul & Linke, Dirk (2019). Differential Adhesion of Trimeric Autotransporter Adhesins to Extracellular Matrix Proteins. Fulltekst i vitenarkiv
• Meuskens, Ina & Linke, Dirk (2019). Binding of Yersinia Ahesin A to ECM molecules: a biosensor approach. Fulltekst i vitenarkiv
• Linke, Dirk & Meuskens, Ina (2018). Adhesion of Yersinia Adhesin A to Collagen. Fulltekst i vitenarkiv
• Meuskens, Ina; Linke, Dirk & Bardiaux, Benjamin (2022). Insights into the Binding Mechanism and Function of Bacterial Adhesins - Interactions between YadA and Host Extracellular Matrix Components. Universitetet i Oslo. Fulltekst i vitenarkiv

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