Gabriele Cordara

• Kersten, Flore; Cortés, Natalia Mojica; Serrano, Albert; Heim, Joel Benjamin; Guyette, Jessica L. & Cezar, Henrique Musseli [Show all 11 contributors for this article] (2026). A Single Amino Acid in the A2 Linker of Cholera Toxin Affects the Efficiency of Toxin Disassembly and Resulting Toxicity. ACS Omega. 11(38), p. 57763–57780. doi: 10.1021/acsomega.6c07022. Full text in Research Archive Show summary

  Abstract Cholera toxin (CT) from Vibrio cholerae and heat-labile enterotoxin (LT) from enterotoxigenic Escherichia coli are highly related AB5-type protein toxins that contain a catalytic A1 subunit, an A2 linker, and a cell-binding homopentamer of B subunits. The two proteins bind the same GM1 receptor and have similar levels of enzymatic activity toward their shared Gsα target, yet CT is more potent than LT. This difference has been mapped to a sequence of 11 amino acids (residues 226–236) near the C-terminus of the A2 linker. We now identify a single amino acid, aspartic acid at position 229, as a key contributor to the function of CT: substituting it for glutamic acid 229 from LT reduced the toxicity of CT to the level of LT. Toxin disassembly by protein disulfide isomerase (PDI), which is an essential event in the intoxication process, was less efficient for CTD229E and LT than for wild-type CT. This was further confirmed by a CT variant, in which four residues were converted into the LT sequence (D229E, I230V, T232I, and H233Y). To test our previous hypothesis that the positioning of the A1 subunit over the B pentamer critically determines toxin disassembly and thus potency, we determined crystal structures of both toxin variants to 1.6 Å resolution and calculated the angle at which the A2 linker exits the pore. The conformations of the two toxin chimeras lie between CT and LT, with the A2 C-terminus of the CT quadruple variant adopting essentially the same fold as LT. The A subunits of the toxin variants as well as that of LT showed a higher conformational freedom than CT as determined by molecular dynamics simulations, suggesting that a more rigid positioning of the A subunit drives toxin disassembly and potency. Our collective data uncover the critical role of D229 in toxin folding and structural dynamics, which in turn impacts toxin potency through effects on the efficiency of PDI-driven toxin disassembly.

• Nguyen, Adam; Heim, Joel Benjamin; Cordara, Gabriele; Chan, Matthew C.; Johannesen, Hedda & Charlesworth, Cristine [Show all 12 contributors for this article] (2026). Shared ligand-blocking mechanism but distinct conformational modulation by α5-targeting antibodies BIIG2 and MINT1526A. Structure. ISSN 0969-2126. doi: 10.1016/j.str.2026.08.001. Full text in Research Archive
• Canals, Mateu Montserrat; Cordara, Gabriele & Krengel, Ute (2025). Allostery. Quarterly Reviews of Biophysics. ISSN 0033-5835. 58(e5), p. 1–26. doi: 10.1017/S0033583524000209. Full text in Research Archive Show summary

  Allostery describes the ability of biological macromolecules to transmit signals spatially through the molecule from an allosteric site – a site that is distinct from orthosteric binding sites of primary, endogenous ligands – to the functional or active site. This review starts with a historical overview and a description of the classical example of allostery – hemoglobin – and other well-known examples (aspartate transcarbamoylase, Lac repressor, kinases, G-protein-coupled receptors, adenosine triphosphate synthase, and chaperonin). We then discuss fringe examples of allostery, including intrinsically disordered proteins and inter-enzyme allostery, and the influence of dynamics, entropy, and conformational ensembles and landscapes on allosteric mechanisms, to capture the essence of the field. Thereafter, we give an overview over central methods for investigating molecular mechanisms, covering experimental techniques as well as simulations and artificial intelligence (AI)-based methods. We conclude with a review of allostery-based drug discovery, with its challenges and opportunities: with the recent advent of AI-based methods, allosteric compounds are set to revolutionize drug discovery and medical treatments.

• Nguyen, Adam; Heim, Joel Benjamin; Cordara, Gabriele; Chan, Matthew C.; Johannesen, Hedda & Charlesworth, Christine [Show all 12 contributors for this article] (2024). Structural and functional characterization of integrin α5-targeting antibodies for anti-angiogenic therapy. BioRxiv. ISSN 0362-4331. doi: 10.1101/2025.01.08.631572. Full text in Research Archive
• Canals, Mateu Montserrat; Bjerregaard-Andersen, Kaare; Sørensen, Henrik Vinther; Kommedal, Eirik; Cordara, Gabriele & Vaaje-Kolstad, Gustav [Show all 7 contributors for this article] (2024). Calcium-binding site in AA10 LPMO from Vibrio cholerae suggests modulating effects during environmental survival and infection. QRB Discovery. 5. doi: 10.1017/qrd.2024.14. Full text in Research Archive Show summary

  Despite major efforts toward its eradication, cholera remains a major health threat and economic burden in many low- and middle-income countries. Between outbreaks, the bacterium responsible for the disease, Vibrio cholerae, survives in aquatic environmental reservoirs, where it commonly forms biofilms, for example, on zooplankton. N-acetyl glucosamine-binding protein A (GbpA) is an adhesin that binds to the chitinaceous surface of zooplankton and breaks its dense crystalline packing thanks to its lytic polysaccharide monooxygenase (LPMO) activity, which provides V. cholerae with nutrients. In addition, GbpA is an important colonization factor associated with bacterial pathogenicity, allowing the binding to mucins in the host intestine. Here, we report the discovery of a cation-binding site in proximity of the GbpA active site, which allows Ca2+, Mg2+, or K+ binding close to its carbohydrate-binding surface. In addition to the X-ray crystal structures of cation-LPMO complexes (to 1.5 Å resolution), we explored how the presence of ions affects the stability and activity of the protein. Calcium and magnesium ions were found to bind to GbpA specifically, with calcium ions – abundant in natural sources of chitin – having the strongest effect on protein stability. When the cation-binding site was rendered non-functional, a decrease in activity was observed, highlighting the importance of the structural elements stabilized by calcium. Our findings suggest a cation-binding site specific to GbpA and related LPMOs that may fine-tune binding and activity for its substrates during environmental survival and host infection.

• Canals, Mateu Montserrat; Bjerregaard-Andersen, Kaare; Sørensen, Henrik Vinther; Cordara, Gabriele; Vaaje-Kolstad, Gustav & Krengel, Ute (2023). Calcium binding site in AA10 LPMO from Vibrio cholerae suggests modulating effects during environment survival and infection. BioRxiv. ISSN 0362-4331. doi: 10.1101/2023.12.22.573012. Full text in Research Archive
• Askarian, Fatemeh; Tsai, Chih-Ming; Cordara, Gabriele; Zurich, Raymond H.; Bjånes, Elisabet & Golten, Ole [Show all 22 contributors for this article] (2023). Immunization with lytic polysaccharide monooxygenase CbpD induces protective immunity against Pseudomonas aeruginosa pneumonia. Proceedings of the National Academy of Sciences of the United States of America (PNAS). ISSN 0027-8424. 120(30). doi: 10.1073/pnas.2301538120. Full text in Research Archive Show summary

  Pseudomonas aeruginosa (PA) CbpD belongs to the lytic polysaccharide monooxygenases (LPMOs), a family of enzymes that cleave chitin or related polysaccharides. Here, we demonstrate a virulence role of CbpD in PA pneumonia linked to impairment of host complement function and opsonophagocytic clearance. Following intratracheal challenge, a PA ΔCbpD mutant was more easily cleared and produced less mortality than the wild-type parent strain. The x-ray crystal structure of the CbpD LPMO domain was solved to subatomic resolution (0.75Å) and its two additional domains modeled by small-angle X-ray scattering and Alphafold2 machine-learning algorithms, allowing structure-based immune epitope mapping. Immunization of naive mice with recombinant CbpD generated high IgG antibody titers that promoted human neutrophil opsonophagocytic killing, neutralized enzymatic activity, and protected against lethal PA pneumonia and sepsis. IgG antibodies generated against full-length CbpD or its noncatalytic M2+CBM73 domains were opsonic and protective, even in previously PA-exposed mice, while antibodies targeting the AA10 domain were not. Preexisting antibodies in PA-colonized cystic fibrosis patients primarily target the CbpD AA10 catalytic domain. Further exploration of LPMO family proteins, present across many clinically important and antibiotic-resistant human pathogens, may yield novel and effective vaccine antigens.

• Cortés, Natalia Mojica; Kersten, Flore; Canals, Mateu Montserrat; III, G. Robb Huhn; Tislevoll, Abelone & Cordara, Gabriele [Show all 8 contributors for this article] (2023). Using Vibrio natriegens for High-Yield Production of Challenging Expression Targets and for Protein Perdeuteration. Biochemistry. ISSN 0006-2960. doi: 10.1021/acs.biochem.3c00612. Full text in Research Archive Show summary

  Production of soluble proteins is essential for structure/function studies; however, this usually requires milligram amounts of protein, which can be difficult to obtain with traditional expression systems. Recently, the Gram-negative bacterium Vibrio natriegens emerged as a novel and alternative host platform for production of proteins in high yields. Here, we used a commercial strain derived from V. natriegens (Vmax X2) to produce soluble bacterial and fungal proteins in milligram scale, which we struggled to achieve in Escherichia coli. These proteins include the cholera toxin (CT) and N-acetyl glucosamine-binding protein A (GbpA) from Vibrio cholerae, the heat-labile enterotoxin (LT) from E. coli and the fungal nematotoxin CCTX2 from Coprinopsis cinerea. CT, GbpA, and LT are secreted by the Type II secretion system in their natural hosts. When these three proteins were produced in Vmax, they were also secreted and could be recovered from the growth media. This simplified the downstream purification procedure and resulted in considerably higher protein yields compared to production in E. coli (6- to 26-fold increase). We also tested Vmax for protein perdeuteration using deuterated minimal media with deuterium oxide as solvent and achieved a 3-fold increase in yield compared to the equivalent protocol in E. coli. This is good news, since isotopic labeling is expensive and often ineffective but represents a necessary prerequisite for some structural biology techniques. Thus, Vmax represents a promising host for production of challenging expression targets and for protein perdeuteration in amounts suitable for structural biology studies.

• Stocker, Christian; Khatanbaatar, Tamjidmaa; Bressan, Luca; Würth-Roderer, Kathrin; Cordara, Gabriele & Krengel, Ute [Show all 7 contributors for this article] (2023). Novel exported fusion enzymes with chorismate mutase and cyclohexadienyl dehydratase activity: Shikimate pathway enzymes teamed up in no man’s land. Journal of Biological Chemistry. ISSN 0021-9258. 299(10). doi: 10.1016/j.jbc.2023.105161. Full text in Research Archive Show summary

  Chorismate mutase (CM) and cyclohexadienyl dehydratase (CDT) catalyze two subsequent reactions in the intracellular biosynthesis of l-phenylalanine (Phe). Here, we report the discovery of novel and extremely rare bifunctional fusion enzymes, consisting of fused CM and CDT domains, which are exported from the cytoplasm. Such enzymes were found in only nine bacterial species belonging to non-pathogenic γ- or β-Proteobacteria. In γ-proteobacterial fusion enzymes, the CM domain is N-terminal to the CDT domain, whereas the order is inverted in β-Proteobacteria. The CM domains share 15% to 20% sequence identity with the AroQγ class CM holotype of Mycobacterium tuberculosis (∗MtCM), and the CDT domains 40% to 60% identity with the exported monofunctional enzyme of Pseudomonas aeruginosa (PheC). In vitro kinetics revealed a Km <7 μM, much lower than for ∗MtCM, whereas kinetic parameters are similar for CDT domains and PheC. There is no feedback inhibition of CM or CDT by the pathway's end product Phe, and no catalytic benefit of the domain fusion compared with engineered single-domain constructs. The fusion enzymes of Aequoribacter fuscus, Janthinobacterium sp. HH01, and Duganella sacchari were crystallized and their structures refined to 1.6, 1.7, and 2.4 Å resolution, respectively. Neither the crystal structures nor the size-exclusion chromatography show evidence for substrate channeling or higher oligomeric structure that could account for the cooperation of CM and CDT active sites. The genetic neighborhood with genes encoding transporter and substrate binding proteins suggests that these exported bifunctional fusion enzymes may participate in signaling systems rather than in the biosynthesis of Phe.

• Sørensen, Henrik Vinther; Canals, Mateu Montserrat; Loose, Jennifer Sarah Maria; Fisher, S. Zoë; Moulin, Martine & Blakeley, Matthew P. [Show all 9 contributors for this article] (2023). Perdeuterated GbpA Enables Neutron Scattering Experiments of a Lytic Polysaccharide Monooxygenase. ACS Omega. 8(32), p. 29101–29112. doi: 10.1021/acsomega.3c02168. Full text in Research Archive Show summary

  Lytic polysaccharide monooxygenases (LPMOs) are surface-active redox enzymes that catalyze the degradation of recalcitrant polysaccharides, making them important tools for energy production from renewable sources. In addition, LPMOs are important virulence factors for fungi, bacteria, and viruses. However, many knowledge gaps still exist regarding their catalytic mechanism and interaction with their insoluble, crystalline substrates. Moreover, conventional structural biology techniques, such as X-ray crystallography, usually do not reveal the protonation state of catalytically important residues. In contrast, neutron crystallography is highly suited to obtain this information, albeit with significant sample volume requirements and challenges associated with hydrogen’s large incoherent scattering signal. We set out to demonstrate the feasibility of neutron-based techniques for LPMOs using N-acetylglucosamine-binding protein A (GbpA) from Vibrio cholerae as a target. GbpA is a multifunctional protein that is secreted by the bacteria to colonize and degrade chitin. We developed an efficient deuteration protocol, which yields >10 mg of pure 97% deuterated protein per liter expression media, which was scaled up further at international facilities. The deuterated protein retains its catalytic activity and structure, as demonstrated by small-angle X-ray and neutron scattering studies of full-length GbpA and X-ray crystal structures of its LPMO domain (to 1.1 Å resolution), setting the stage for neutron scattering experiments with its substrate chitin.

• Thorbjørnsrud, Helen Vikdal; Bressan, Lucas; Khatanbaatar, Tamjidmaa; Carrer, Manuel; Würth-Roderer, Kathrin & Cordara, Gabriele [Show all 9 contributors for this article] (2023). What Drives Chorismate Mutase to Top Performance? Insights from a Combined In Silico and In Vitro Study. Biochemistry. ISSN 0006-2960. 62(3), p. 782–796. doi: 10.1021/acs.biochem.2c00635. Full text in Research Archive Show summary

  Unlike typical chorismate mutases, the enzyme from Mycobacterium tuberculosis (MtCM) has only low activity on its own. Remarkably, its catalytic efficiency kcat/Km can be boosted more than 100-fold by complex formation with a partner enzyme. Recently, an autonomously fully active MtCM variant was generated using directed evolution, and its structure was solved by X-ray crystallography. However, key residues were involved in crystal contacts, challenging the functional interpretation of the structural changes. Here, we address these challenges by microsecond molecular dynamics simulations, followed up by additional kinetic and structural analyses of selected sets of specifically engineered enzyme variants. A comparison of wild-type MtCM with naturally and artificially activated MtCMs revealed the overall dynamic profiles of these enzymes as well as key interactions between the C-terminus and the active site loop. In the artificially evolved variant of this model enzyme, this loop is preorganized and stabilized by Pro52 and Asp55, two highly conserved residues in typical, highly active chorismate mutases. Asp55 stretches across the active site and helps to appropriately position active site residues Arg18 and Arg46 for catalysis. The role of Asp55 can be taken over by another acidic residue, if introduced at position 88 close to the C-terminus of MtCM, as suggested by molecular dynamics simulations and confirmed by kinetic investigations of engineered variants.

• Thorbjørnsrud, Helen Vikdal; Bressan, Luca; Khatanbaatar, Tamjidmaa; Carrer, Manuel; Würth-Roderer, Kathrin & Cordara, Gabriele [Show all 9 contributors for this article] (2022). What drives chorismate mutase to top performance? Insights from a combined in silico and in vitro study. BioRxiv. ISSN 0362-4331. doi: 10.1101/2022.11.08.515563. Full text in Research Archive
• Skåne, Anna; Edvardsen, Per Kristian Thorén; Cordara, Gabriele; Loose, Jennifer Sarah Maria; Leitl, Kira & Krengel, Ute [Show all 9 contributors for this article] (2022). Chitinolytic enzymes contribute to the pathogenicity of Aliivibrio salmonicida LFI1238 in the invasive phase of cold-water vibriosis. BMC Microbiology. 22(1). doi: 10.1186/s12866-022-02590-2. Full text in Research Archive Show summary

  Background - Aliivibrio salmonicida is the causative agent of cold-water vibriosis in salmonids (Oncorhynchus mykiss and Salmo salar L.) and gadidae (Gadus morhua L.). Virulence-associated factors that are essential for the full spectrum of A. salmonicida pathogenicity are largely unknown. Chitin-active lytic polysaccharide monooxygenases (LPMOs) have been indicated to play roles in both chitin degradation and virulence in a variety of pathogenic bacteria but are largely unexplored in this context.<p> <p>Results - In the present study we investigated the role of LPMOs in the pathogenicity of A. salmonicida LFI238 in Atlantic salmon (Salmo salar L.). In vivo challenge experiments using isogenic deletion mutants of the two LPMOs encoding genes AsLPMO10A and AsLPMO10B, showed that both LPMOs, and in particular AsLPMO10B, were important in the invasive phase of cold-water vibriosis. Crystallographic analysis of the AsLPMO10B AA10 LPMO domain (to 1.4 Å resolution) revealed high structural similarity to viral fusolin, an LPMO known to enhance the virulence of insecticidal agents. Finally, exposure to Atlantic salmon serum resulted in substantial proteome re-organization of the A. salmonicida LPMO deletion variants compared to the wild type strain, indicating the struggle of the bacterium to adapt to the host immune components in the absence of the LPMOs.<p> <p>Conclusion - The present study consolidates the role of LPMOs in virulence and demonstrates that such enzymes may have more than one function.

• Manna, Dipankar; Cordara, Gabriele & Krengel, Ute (2020). Crystal structure of MOA in complex with a peptide fragment: A protease caught in flagranti. Current Research in Structural Biology. ISSN 2665-928X. doi: 10.1016/j.crstbi.2020.04.003. Full text in Research Archive Show summary

  The Marasmius oreades agglutinin (MOA) is the holotype of an emerging family of fungal chimerolectins and an active Ca2+/Mn2+-dependent protease, which exhibits a unique papain-like fold with special active site features. Here we investigated the functional significance of the structural elements differentiating MOA from other papain-like cysteine proteases. X-ray crystal structures of MOA co-crystallized with two synthetic substrates reveal cleaved peptides bound to the catalytic site, corresponding to the final products of the proteolytic reaction. Anomalous diffraction data on crystals grown in the presence of calcium and manganese, cadmium or zinc resolve the calcium/manganese preference of MOA and elucidate the inhibitory roles of zinc and cadmium towards papain-like cysteine proteases in general. The reported structures, together with activity data of MOA active site variants, point to a conservation of the general proteolysis mechanism established for papain. Ultimately, the findings suggest that papain and the papain-like domain of MOA are the product of convergent evolution.

• Cordara, Gabriele; Bjerregaard-Andersen, Kaare & Krengel, Ute (2018). Virus-like immune defense protein in mushrooms. Structure. ISSN 0969-2126. 26(3), p. 369–371. doi: 10.1016/j.str.2018.02.013. Full text in Research Archive Show summary

  Fungi and plants do not have an adaptive immune system. Innate immunity serves as their sole defense, often based on carbohydrate recognition by lectins. In a twist of nature, as revealed by Sommer et al. (2018) in this issue of Structure, a conserved fungal immunoprotein adopts the shape of a miniature virus.

• Cordara, Gabriele; Manna, Dipankar & Krengel, Ute (2017). A family of papain-like fungal chimerolectins with distinct Ca2+-dependent activation mechanism. Biochemistry. ISSN 0006-2960. 56(35), p. 4689–4700. doi: 10.1021/acs.biochem.7b00317. Full text in Research Archive Show summary

  An important function of fungal lectins is to protect their host. Marasmius oreades agglutinin (MOA) is toxic to nematodes and exerts its protective effect through protease activity. Its proteolytic function is associated with a papain-like dimerization domain. The closest homologue of MOA is Polyporus squamosus lectin 1a (PSL1a). Here, we probed PSL1a for catalytic activity and confirmed that it is a calcium-dependent cysteine protease, like MOA. The X-ray crystal structures of PSL1a (1.5 Å) and MOA (1.3 Å) in complex with calcium and the irreversible cysteine protease inhibitor E-64 elucidated the structural basis for their mechanism of action. The comparison with other calcium-dependent proteases (calpains, LapG) reveals a unique metal-dependent activation mechanism relying on a calcium-induced backbone shift and intradimer cooperation. Intriguingly, the enzymes appear to use a tyrosine-gating mechanism instead of pro-peptide processing. A search for potential MOA orthologues suggests the existence of a whole new family of fungal chimerolectins with these unique features.

• Manna, Dipankar; Pust, Sascha; Torgersen, Maria Lyngaas; Cordara, Gabriele; Kunzler, Markus & Krengel, Ute [Show all 7 contributors for this article] (2017). Polyporus squamosus lectin 1a (PSL1a) exhibits cytotoxicity in mammalian cells by disruption of focal adhesions, inhibition of protein synthesis and induction of apoptosis. PLOS ONE. 12(1). doi: 10.1371/journal.pone.0170716. Full text in Research Archive Show summary

  PSL1a is a lectin from the mushroom Polyporus squamosus that binds to sialylated glycans and glycoconjugates with high specificity and selectivity. In addition to its N-terminal carbohydrate-binding domain, PSL1a possesses a Ca2+-dependent proteolytic activity in the C-terminal domain. In the present study, we demonstrate that PSL1a has cytotoxic effects on mammalian cancer cells, and we show that the cytotoxicity is dependent on the cysteine protease activity. PSL1a treatment leads to cell rounding and detachment from the substratum, concomitant with disruption of vinculin complexes in focal adhesions. We also demonstrate that PSL1a inhibits protein synthesis and induces apoptosis in HeLa cells, in a time- and concentration-dependent manner.

• Cordara, Gabriele; Eerde, Jan Hendrik Van; Grahn, Elin; Winter, HC; Goldstein, IJ & Krengel, Ute (2016). An unusual member of the papain superfamily: Mapping the catalytic cleft of the Marasmius oreades agglutinin (MOA) with a caspase inhibitor. PLOS ONE. 11(2). doi: 10.1371/journal.pone.0149407. Full text in Research Archive Show summary

  Papain-like cysteine proteases (PLCPs) constitute the largest group of thiol-based protein degrading enzymes and are characterized by a highly conserved fold. They are found in bacteria, viruses, plants and animals and involved in a number of physiological and pathological processes, parasitic infections and host defense, making them interesting targets for drug design. The Marasmius oreades agglutinin (MOA) is a blood group B-specific fungal chimerolectin with calcium-dependent proteolytic activity. The proteolytic domain of MOA presents a unique structural arrangement, yet mimicking the main structural elements in known PLCPs. Here we present the X-ray crystal structure of MOA in complex with Z-VAD-fmk, an irreversible caspase inhibitor known to cross-react with PLCPs. The structural data allow modeling of the substrate binding geometry and mapping of the fundamental enzyme-substrate interactions. The new information consolidates MOA as a new, yet strongly atypical member of the papain superfamily. The reported complex is the first published structure of a PLCP in complex with the well characterized caspase inhibitor Z-VAD-fmk.

• Cordara, Gabriele; Winter, Harry C.; Goldstein, Irwin J.; Krengel, Ute & Sandvig, Kirsten (2014). The fungal chimerolectin MOA inhibits protein and DNA synthesis in NIH/3T3 cells and may induce BAX-mediated apoptosis. Biochemical and Biophysical Research Communications - BBRC. ISSN 0006-291X. 447(4), p. 586–589. doi: 10.1016/j.bbrc.2014.04.043. Full text in Research Archive
• Cordara, Gabriele & Krengel, Ute (2012). Structure determination of lectins by x-ray crystallography - a hands-on approach. In Rauter, A.P. & Lindhorst, T.K. (Ed.), Carbohydrate Chemistry - Chemical and Biological Approaches. Royal Society of Chemistry (RSC). ISSN 9781849731546. p. 222–246. doi: 10.1039/9781849737173-00222. Full text in Research Archive
• Cordara, Gabriele; Jacobsen, Wolfgang Mathias; Johansen, Harald Thidemann; Winter, Harry C.; Goldstein, IJ & Sandvig, Kirsten [Show all 7 contributors for this article] (2011). Marasmius oreades agglutinin (MOA) is a chimerolectin with proteolytic activity. Biochemical and Biophysical Research Communications - BBRC. ISSN 0006-291X. 408(3), p. 405–410. doi: 10.1016/j.bbrc.2011.04.031. Full text in Research Archive

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• Cortés, Natalia Mojica; Kersten, Flore; Serrano, Albert; Heim, Joel Benjamin; Cordara, Gabriele & Teter, Ken [Show all 8 contributors for this article] (2026). Why is cholera toxin more toxic than heat-labile enterotoxin? Insights from protein crystallography and MD simulations. Full text in Research Archive
• Albrechtova, Ema; Cordara, Gabriele & Krengel, Ute (2024). Structural characterisation of ATP-induced disassembly mechanisms in cytolethal distending toxin. Full text in Research Archive
• Cortés, Natalia Mojica; Kersten, Flore; Krengel, Ute; Teter, Ken; Cordara, Gabriele & Heim, Joel Benjamin [Show all 7 contributors for this article] (2024). Molecular insights into the differential toxicity of cholera toxin and heat-labile enterotoxin. Full text in Research Archive
• Cortés, Natalia Mojica; Kersten, Flore; Krengel, Ute; Cordara, Gabriele; Heim, Joel Benjamin & Teter, Ken [Show all 7 contributors for this article] (2024). Structural basis for the differential toxicity of cholera toxin and heat-labile enterotoxin. Full text in Research Archive
• Cortés, Natalia Mojica; Kersten, Flore; Krengel, Ute; Cordara, Gabriele; Serrano, Albert & Heim, Joel Benjamin [Show all 7 contributors for this article] (2024). Molecular insights into the differential toxicity of cholera toxin and heat-labile enterotoxin. Full text in Research Archive
• Kersten, Flore; Cordara, Gabriele & Krengel, Ute (2024). Cryo-EM structure of a fungal chimerolectin. Full text in Research Archive
• Canals, Mateu Montserrat; Bjerregaard-Andersen, Kaare; Sørensen, Henrik Vinther; Kommedal, Eirik; Cordara, Gabriele & Vaaje-Kolstad, Gustav [Show all 7 contributors for this article] (2024). Calcium binding site in AA10 LPMO from Vibrio cholerae suggests modulating effects during environmental survival and infection                                               . Full text in Research Archive
• Stocker, Christian; Khatanbaatar, Tamjidmaa; Bressan, Luca; Würth-Roderer, Kathrin; Cordara, Gabriele & Krengel, Ute [Show all 7 contributors for this article] (2023). Novel exported bifunctional fusion enzymes with chorismate mutase and cyclohexadienyl dehydratase activity: shikimate pathway enzymes teamed up in no man's land. BioRxiv. ISSN 0362-4331. doi: 10.1101/2023.03.13.532365. Full text in Research Archive
• Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute (2023). Cryo-EM reveals a novel domain fold of a fungal toxin. Full text in Research Archive
• Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute (2022). Cryo-EM structure of a fungal chimerolectin. Full text in Research Archive
• Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute (2022). Cryo-EM structure of a fungal toxin. Full text in Research Archive
• Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute (2022). Cryo-EM structure of a fungal toxin. Full text in Research Archive
• Manna, Dipankar; Cordara, Gabriele; Sandvig, Kirsten & Krengel, Ute (2016). I Robot- Biological nanomachines for fungal defense. Full text in Research Archive
• Manna, Dipankar; Cordara, Gabriele; Pust, Sascha; Goldstein, Irwin J; Winter, Harry C. & Krengel, Ute [Show all 7 contributors for this article] (2015). PSL1a: a new member in the growing family of fungal chimerolectin. Full text in Research Archive
• Manna, Dipankar; Cordara, Gabriele; Goldstein, Irwin J. & Krengel, Ute (2014). Conserved catalytic machinery in the MOA-PSL family of fungal chimerolectins. Full text in Research Archive
• Manna, Dipankar; Cordara, Gabriele; Goldstein, Irwin J. & Krengel, Ute (2013). Towards structural and functional characterization of the Marasmius oreades agglutinin (MOA) and its role in host-pathogen interactions. Full text in Research Archive
• Cordara, Gabriele (2013). Thermofluor - A complementary technique in structural biology. Full text in Research Archive
• Cordara, Gabriele (2013). Structure-function studies on the proteolytic mechanism of the Marasmius oreades agglutinin (MOA), a cytotoxic lectin. Full text in Research Archive
• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, Irwin; Sandvig, Kirsten & Krengel, Ute (2012). Marasmius oreades Agglutinin (MOA): proteolytic activity of a cytotoxic lectin. Full text in Research Archive Show summary

  Lectins, sugar binding proteins of non-immune origin, are one of the many weapons used by plants and fungi to defend themselves against potential predators and parasites. This role is often fulfilled by the presence of an additional domain or subunit with catalytic function. The Marasmius oreades agglutinin (MOA) is a Gal-α3Gal specific lectin extracted from the fruiting body of the Marasmius oreades mushroom. Parenterally administered MOA triggers a systemic effect in mice which closely resembles the symptoms of the shiga toxin-induced hemolytic uremic syndrome (Stx-HUS) in humans. The structure of MOA in complex calcium, recently solved by X-ray crystallography, elucidated the fine details of the sugar binding specificity of the N-terminal ricin B chain-like domain and showed the existence of a C-terminal domain with an α/β hydrolase fold. A search for structural homologues on the DALI database, using the coordinates of the C-terminal domain as the input, provided a significant number of top-ranking hits belonging to the family of the papain-like cysteine peptidases (clan CA). A notable feature is the conservation in MOA of two key residues of the catalytic triad (Cys215 and His257) while the third residue is substituted by an amino acid with similar properties (Glu274 in place of Asp), suggesting a potential hydrolytic activity associated to the C-terminal domain of MOA. Starting from this structure-based functional hypothesis, MOA was probed with a model substrate and found to be a calcium-dependent cysteine peptidase, with Cys215 acting as the catalytic residue.

• Munack, Steffi; Roderer, Kathrin; Kamarauskaite, Jurate; Cordara, Gabriele; Eerde, André van & Kast, Peter [Show all 7 contributors for this article] (2012). Towards New Drugs Against Tuberculosis. Full text in Research Archive
• Cordara, Gabriele; Goldstein, Irwin; Sandvig, Kirsten & Krengel, Ute (2012). Structure-function studies on the proteolytic activity of the Marasmius oreades agglutinin (MOA), a cytotoxic lectin. Full text in Research Archive Show summary

  Lectins, sugar binding proteins of non-immune origin, are one of the many weapons used by plants and fungi to defend themselves against potential predators and parasites. This role is often fulfilled by the presence of an additional domain or subunit with catalytic function. The Marasmius oreades agglutinin (MOA) is a Gal-3Gal specific lectin extracted from the fruiting body of the Marasmius oreades mushroom. Parenterally administered MOA triggers a systemic effect in mice, which closely resembles the symptoms of the shiga toxin-induced hemolytic uremic syndrome (Stx-HUS) in humans. The structure of the MOA homodimer in complex with calcium, solved by X-ray crystallography [1], suggests an enzymatic function associated to the C-terminal dimerization domain, experimentally confirmed to be associated with proteolytic activity [2]. The enzymatically active domain of MOA shows an / hydrolase fold, bearing a structural resemblance to the enzymes of the papain-like cysteine peptidase family (clan CA). A notable feature in MOA is the conservation of two key residues of the catalytic triad (Cys215 and His257), while the third residue is substituted by an amino acid with similar properties (Glu274 in place of Asp). Here, we present further evidence, including crystallographic data, pointing to a key role of these residues for catalysis. [1] Grahn, E.M., Winter, H.C., Tateno, H., Goldstein, I.J., Krengel, U., (2009). J. Mol.. Biol., 390, 457–466 [2] Cordara, G., Egge-Jacobsen, W., Johansen, H.T., Winter, H.C., Goldstein, I.J., Sandvig, K., Krengel, U. (2011). Biochem. Biophys. Res. Commun., 408, 405-410.

• Munack, Steffi; Cordara, Gabriele; Eerde, Jan Hendrik Van; Kamarauskaite, Jurate; Roderer, Kathrin & Kast, Peter [Show all 7 contributors for this article] (2012). Towards New Drugs Against Tuberculosis. Full text in Research Archive
• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, Irwin; Sandvig, Kirsten & Krengel, Ute (2011). MOA : structure-driven discovery of the proteolytic activity of a cytotoxic lectin. Full text in Research Archive Show summary

  Lectins, sugar binding proteins of non-immune origin, are one of the many weapons used by plants and fungi to defend themselves against potential predators and parasites. This role is often fulfilled by the presence of a domain with an additional function besides the sugar binding domain. The Marasmius oreades agglutinin (MOA) is a Gal-α3Gal specific lectin extracted from the fruiting body of the Marasmius oreades mushroom. Other than showing the rare ability to specifically agglutinate erythrocytes of the blood group B (containing the recognized disaccharide on the non-reducing end of the B glycotope), parenterally administered MOA triggers a systemic effect in mice which closely resembles the symptoms of the shiga toxin-induced hemolytic uremic syndrome (Stx-HUS) in humans. The structure of MOA in complex with the branched blood group B trisaccharide and calcium, recently solved by X-ray crystallography, elucidated the fine details of the sugar binding specificity of the N-terminal ricin B chain-like domain and showed the existence of an alpha/beta hydrolase fold C-terminal domain involved in dimerization A search for structural homologues, using the coordinates of the C-terminal domain as the input for the DALI database, provided a significant number of top-ranking hits belonging to the family of the papain-like cysteine peptidates (clan CA). A notable feature is the conservation in MOA of two key residues of the catalytic triad (Cys215 and His257) while the third residue is substituted by an amino acid with similar properties (Glu274 in place of Asp). Starting from the structure-based functional hypothesis of the presence of a hydrolytic activity associated to the C-terminal domain, MOA was probed with a model substrate and found to be a calcium-dependent cysteine peptidase, with Cys215 acting as the catalytic residue.

• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, IJ; Sandvig, Kirsten & Krengel, Ute (2011). Marasmius oreades agglutinin (MOA) is a cytotoxic lectin with proteolytic activity. Full text in Research Archive Show summary

  The Marasmius oreades mushroom lectin (MOA) is known since the 1950s for its blood group type B binding specificity. The three-dimensional structure of the lectin, recently determined by X-ray crystallography elucidated the details of its carbohydrate binding characteristics, and in particular how this lectin discriminates between blood group A and B structures. MOA’s C-terminal domain adopts an alpha/beta fold. This domain primarily serves as the dimerization interface for MOA protomers, although its structural similarity to hydrolytic enzymes suggests that it may exhibit additional functions. Here, we show that MOA indeed has a previously unknown proteolytic activity linked to its C-terminal domain, involving Cys215 as the catalytically active residue and a strict dependence on the calcium ion fulfilling a fundamental structural role.

• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, IJ; Sandvig, Kirsten & Krengel, Ute (2011). Marasmius oreades agglutinin (MOA): A lectin with a proteolytic domain. Full text in Research Archive
• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, IJ; Sandvig, Kirsten & Krengel, Ute (2011). MOA : structure-driven discovery of the proteolytic activity of a cytotoxic lectin. Full text in Research Archive Show summary

  The Hemolytic Uremic Syndrome (HUS) is a disease characterized by hemolytic anemia, low platelet count and renal impairment. Based on the causative agent HUS can be classified as Shiga toxin-associated HUS (Stx-HUS) or non-Shiga toxin associated HUS (non-Stx-HUS), the former accounting for 70% of HUS cases in Europe and the North America. The therapy for HUS is still merely supportive, and there is currently no viable animal model (besides baboons) for a more in-depth systemic study of the disease. The Marasmius oreades agglutinin (MOA) is a two-domains Galα3Gal specific lectin, a sugar binding protein of non-immune origin, extracted from the fruiting body of the Marasmius oreades mushrooms. Other than showing the rare ability to specifically agglutinate erythrocytes of the blood group B, MOA parenterally administered to mice triggers a systemic effect which closely resembles the symptoms of Stx-HUS in humans. The recently determined x-ray crystallographic structure of MOA in complex with calcium and the blood group B branched trisaccharide (PDB id 2IHO) confirm the existence of a ricin B chain-like sugar binding domain and shows a C-terminal alpha/beta folded domain. A search for structural homologues on the DALI database using the coordinates of the C-terminal domain suggests a possible enzymatic function for the non-sugar binding domain, thus providing a further link to the Shiga toxin induced HUS. The investigation of the enzymatic activity of MOA has confirmed the structure-based functional hypothesis and it has shown that the protein behaves as a Ca2+-dependent cysteine protease. An initial characterization of the enzymatic activity has been performed, showing how the processing of the substrate is affected by the concentration of Ca2+, the presence of a reducing agent and other factors. At the same time, the cytotoxic activity of MOA on different cell types has been evaluated, showing the NIH3T3 cell line as the most sensitive among the tested targets; the incubation with MOA leads to cell detachment, preceded by a decrease in the level of protein and DNA synthesis.

• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, IJ; Sandvig, Kirsten & Krengel, Ute (2011). MOA : structure-driven discovery of the proteolytic activity of a cytotoxic lectin. Full text in Research Archive Show summary

  Lectins, sugar binding proteins of non-immune origin, are one of the many weapons used by plants and fungi to defend themselves against potential predators and parasites. This role is often fulfilled by the presence of a domain with an additional function besides the sugar binding domain. The Marasmius oreades agglutinin (MOA) is a Gal-alpha3Gal specific lectin extracted from the fruiting body of the Marasmius oreades mushroom. Other than showing the rare ability to specifically agglutinate erythrocytes of the blood group B (containing the recognized disaccharide on the non-reducing end of the B glycotope), parenterally administered MOA triggers a systemic effect in mice which closely resembles the symptoms of the shiga toxin-induced hemolytic uremic syndrome (Stx-HUS) in humans. The structure of MOA in complex with the branched blood group B trisaccharide and calcium, recently solved by X-ray crystallography, elucidated the fine details of the sugar binding specificity of the N-terminal ricin B chain-like domain and showed the existence of an alpha/beta hydrolase fold C-terminal domain involved in dimerization A search for structural homologues, using the coordinates of the C-terminal domain as the input for the DALI database, provided a significant number of top-ranking hits belonging to the family of the papain-like cysteine peptidates (clan CA). A notable feature is the conservation in MOA of two key residues of the catalytic triad (Cys215 and His257) while the third residue is substituted by an amino acid with similar properties (Glu274 in place of Asp). Starting from the structure-based functional hypothesis of the presence of a hydrolytic activity associated to the C-terminal domain, MOA was probed with a model substrate and found to be a calcium-dependent cysteine peptidase, with Cys215 acting as the catalytic residue.

• Cordara, Gabriele; Grahn, Elin; Jacobsen, Wolfgang Mathias; Goldstein, IJ; Sandvig, Kirsten & Krengel, Ute (2010). MOA – A cytotoxic lectin from a mushroom could provide an animal model for HUS/TPP. Full text in Research Archive Show summary

  The Hemolytic Uremic Syndrome (HUS) is a disease characterized by hemolytic anemia, low platelet count and renal impairment. Based on the causative agent HUS can be classified as Shiga toxin-associated HUS (Stx-HUS) or non-Shiga toxin associated HUS (non-Stx-HUS), the former accounting for 70% of HUS cases in Europe and the North America. The therapy for HUS is still merely supportive, and there is currently no viable animal model (aside baboons) for a more in-depth systemic study of the disease. The Marasmius Oreades Agglutinin (MOA) is a two-domains Gal-alpha1,3Gal specific lectin, a sugar binding protein of non-immune origin, extracted from the fruiting body of the Marasmius oreades mushrooms. Other than showing the rare ability to specifically agglutinate erythrocytes of the blood group B, parenterally administered MOA triggers a systemic effect in mice which closely resembles the symptoms of Stx-HUS in humans. The recently determined x-ray crystallographic structure of MOA in complex with calcium and the blood group B branched trisaccharide suggests a possible enzymatic function for the non-sugar binding domain, thus providing a further link to the Shiga toxin induced HUS. The aim of this project is to investigate the nature of MOA’s enzymatic function along with structurally relevant determinant involved in the enzymatic activity and to determine the relationship between such activity and the cytotoxic effects.

• Cordara, Gabriele (2010). MOA - s sugar binding protein - From structure to hypotheses on function. Full text in Research Archive
• Cordara, Gabriele; Grahn, Elin; Goldstein, IJ & Krengel, Ute (2010). MOA, a sugar binding protein - From structure to hypotheses on function. Full text in Research Archive
• Cordara, Gabriele; Krengel, Ute & Sandvig, Kirsten (2013). Structure-function studies on the Marasmius oreades agglutinin (MOA), a cytotoxic protease. Akademika Publishing. Full text in Research Archive

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