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Montserrat Canals, Mateu; Sekulic, Nikolina & Krengel, Ute
(2024).
UiO structural biology core facilities.
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Krengel, Ute
(2023).
Protein-Carbohydrate Interactions in Infection and Cancer Biology.
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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 2692-8205.
doi:
10.1101/2023.03.13.532365.
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Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute
(2023).
Cryo-EM reveals a novel domain fold of a fungal toxin.
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Montserrat Canals, Mateu; Sørensen, Henrik Vinther; Oksanen, Esko & Krengel, Ute
(2022).
Neutron studies of the bacterial colonization factor GbpA.
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Montserrat Canals, Mateu; Sørensen, Henrik Vinther; Oksanen, Esko O. & Krengel, Ute
(2022).
Structural studies on GbpA using neutron scattering methods.
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Montserrat Canals, Mateu; Bjerregaard-Andersen, Kaare; Sørensen, Henrik Vinther & Krengel, Ute
(2022).
How salts modulate activity in chitin-active LPMOs relevant for environmental survival and human pahtogenicity.
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Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute
(2022).
Cryo-EM structure of a fungal toxin.
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Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute
(2022).
Cryo-EM structure of a fungal toxin.
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Kersten, Flore Marie Colette; Cordara, Gabriele & Krengel, Ute
(2022).
Cryo-EM structure of a fungal chimerolectin.
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Sekulic, Nikolina & Krengel, Ute
(2022).
PX-Oslo: Structural Biology Core Facilities
.
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Bousquet, Paula; Sandvik, Joe Alexander; Edin, Nina Frederike Jeppesen & Krengel, Ute
(2022).
Corrigendum to: Hypothesis: Hypoxia induces de novo synthesis of NeuGc gangliosides in humans through CMAH domain substitute (Biochemical and Biophysical Research Communications (2018) 495(1) (1562–1566), (S0006291X17323616), (10.1016/j.bbrc.2017.11.183)).
Biochemical and Biophysical Research Communications - BBRC.
ISSN 0006-291X.
587.
doi:
10.1016/j.bbrc.2021.11.074.
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Krengel, Ute
(2022).
Academia meets Industry.
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Krengel, Ute
(2022).
Molecular Mechanisms of Cholera Pathogenesis.
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Krengel, Ute
(2021).
My experience with HALOS: Making the most of structural biology
.
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Mojica Cortez, Natalia & Krengel, Ute
(2020).
PX-Oslo- UiO Structural Biology Core Facilities.
[Internet].
Department of Chemistry, UiO.
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Pesci, Giacomo; Johannesen, Hedda; Løset, Geir Åge & Krengel, Ute
(2020).
A promising antibody to treat cancer. How does it work?
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Sørensen, Henrik Vinther; Bjerregaard-Andersen, Kaare; Vaaje-Kolstad, Gustav; Krengel, Ute & Lund, Reidar
(2020).
Applying soft matter techniques to understand bacterial colonization.
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Johannesen, Hedda; Bjerregaard-Andersen, Kaare; Grzybek, Michal; Løset, Geir Åge & Krengel, Ute
(2020).
Molecular basis of specificity of a promising anti-tumour antibody .
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Krengel, Ute
(2020).
UiO Structural Biology Core Facilities: Who Are We and How Can We Help You?
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Mojica Cortez, Natalia; Sekulic, Nikolina & Krengel, Ute
(2020).
UiO Structural Biology Core Facilities: Who Are We and How Can We Help You?
.
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Krengel, Ute
(2019).
Structural Chemistry - Past, Presence and Future.
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Heggelund, Julie Elisabeth; Heim, Joel Benjamin & Krengel, Ute
(2019).
Cholera blood group association elucidated with synchrotron radiation.
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Johannesen, Hedda; Bjerregaard-Andersen, Kaare; Løset, Geir Åge & Krengel, Ute
(2019).
An anti-tumour antibody that specifically recognises N-glycolyl GM3.
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Johannesen, Hedda; Bjerregaard-Andersen, Kaare; Grzybek, Michal; Ünal, Coskun; Løset, Geir Åge & Krengel, Ute
(2019).
Crystal structure of an anti-tumour antibody in complex with its tumour-specific antigen.
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Sørensen, Henrik Vinther; Krengel, Ute; Lund, Reidar; Browning, Kathryn & Bjerregaard-Andersen, Kaare
(2019).
Bacterial colonization investigated with X-ray and neutron techniques.
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Sørensen, Henrik Vinther; Krengel, Ute; Vaaje-Kolstad, Gustav; Lund, Reidar; Browning, Kathryn & Bjerregaard-Andersen, Kaare
[Show all 7 contributors for this article]
(2019).
Using X-rays and neutrons to understand a bacterial
colonization factor.
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Sørensen, Henrik Vinther; Krengel, Ute; Browning, Kathryn; Bjerregaard-Andersen, Kaare; Cardenas, Marite & Lund, Reidar
[Show all 7 contributors for this article]
(2019).
Study of bacterial colonization with X-ray and neutron scattering techniques.
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Johannesen, Hedda; Bjerregaard-Andersen, Kaare; Heggelund, Julie Elisabeth; Løset, Geir Åge & Krengel, Ute
(2019).
14F7 - A tumour specific antibody
.
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Sørensen, Henrik Vinther; Browning, Kathryn; Bjerregaard-Andersen, Kaare; Lund, Reidar & Krengel, Ute
(2019).
Applying X-rays and neutrons to understand a bacterial virulence factor.
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Johannesen, Hedda; Bjerregaard-Andersen, Kaare; Heggelund, Julie Elisabeth; Løset, Geir Åge & Krengel, Ute
(2018).
NeuGc GM3 - a cancer-specific ganglioside that is recognised by the antibody 14F7.
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Sørensen, Henrik Vinther; Browning, Kathryn; Bjerregaard-Andersen, Kaare; Lund, Reidar & Krengel, Ute
(2018).
Borrowing material science techniques to study biological problems.
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Johannesen, Hedda; Mykland hoås, Helene; Heggelund, Julie Elisabeth; Bjerregaard-Andersen, Kaare; Løset, Geir Åge & Krengel, Ute
(2018).
14F7, An anti-ganglioside scFv for cancer therapies.
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Johannesen, Hedda; Mykland Hoås, Helene; Heggelund, Julie Elisabeth; Bjerregaard-Andersen, Kaare; Løset, Geir Åge & Krengel, Ute
(2018).
14F7, an antibody that recognises a tumour-specific ganglioside with high specificity
.
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Krengel, Ute
(2018).
Cholera Toxin: Molecular Mechanisms and Drug Design
.
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Krengel, Ute
(2018).
Remote Control by Inter-Enzyme Allostery -
Or: The Marvellous World of Chorismate Mutases
.
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Krengel, Ute
(2018).
Structure-Function Studies of Cholera Toxin: Molecular Underpinnings of Human Disease.
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Krengel, Ute
(2018).
Interesting examples of allostery (?).
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Krengel, Ute
(2018).
How to design posters and write competitive grant proposals.
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Sørensen, Henrik Vinther; Bjerregaard-Andersen, Kaare; Lund, Reidar & Krengel, Ute
(2018).
A bacterial colonization factor studied by X-ray and neutron techniques.
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Krengel, Ute
(2018).
Protein crystallography: Glimpses from my career.
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Sørensen, Henrik Vinther; Browning, Kathryn; Lund, Reidar; Bjerregaard-Andersen, Kaare & Krengel, Ute
(2018).
A surface active enzyme and colonization factor investigated by x-ray and neutron techniques.
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Krengel, Ute
(2018).
How to become an academic leader?
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Heim, Joel Benjamin; Teter, Kenneth & Krengel, Ute
(2018).
Towards the molecular basis for the differential toxicity of cholera toxin and heat-labile enterotoxin.
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Johannesen, Hedda; Hoås Mykland, Helene; Heggelund, Julie Elisabeth; Bjerregaard-Andersen, Kaare; Løset, Geir Åge & Krengel, Ute
(2018).
An anti-ganglioside scFv for cancer therapies.
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Sørensen, Henrik Vinther; L. Browning, Kathryn; Lund, Reidar; Krengel, Ute & Bjerregaard-Andersen, Kaare
(2018).
A surface active enzyme and colonization factor investigated by x-ray and neutron scattering.
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Sørensen, Henrik Vinther; Browning, Kathryn; Lund, Reidar; Bjerregaard-Andersen, Kaare & Krengel, Ute
(2017).
A surface active enzyme and colonization factor investigated by x-ray and neutron scattering - Hercules.
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Krengel, Ute
(2017).
Molecular Mechanisms of Disease.
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Thorbjørnsrud, Helen Vikdal; Fahrig-Kamarauskaite, Jurate; Burschowsky, Daniel; Kast, Peter & Krengel, Ute
(2017).
Structural elucidation of highly active chorismate mutase variants obtained by directed evolution.
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Bjerregaard-Andersen, Kaare; Harby, Caroline; Loose, Jennifer Sarah Maria; Vaaje-Kolstad, Gustav & Krengel, Ute
(2017).
Chasing protons in lytic polysaccharide mono oxygenases - studying the Vibrio colonization factor GbpA by neutron crystallography.
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Bjerregaard-Andersen, Kaare; Harby, Caroline & Krengel, Ute
(2017).
TOWARDS NEW INSIGHTS INTO VIBRIO CHOLERAE COLONIZATION BY X-RAY AND NEUTRON DIFFRACTION.
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Heim, Joël Benjamin; Teter, Kenneth & Krengel, Ute
(2017).
Molecular interactions of cholera toxin – working with unstable proteins and complexes.
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Tveitan, Siri Luise; Heim, Joël Benjamin & Krengel, Ute
(2017).
Structural Studies on Cholera.
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Heim, Joël Benjamin; Teter, Kenneth & Krengel, Ute
(2017).
Molecular interactions of cholera toxin – working with unstable proteins and complexes.
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Krengel, Ute
(2017).
Molecular Basis of Blood-Group-Dependent Diseases Like Cholera.
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Krengel, Ute
(2017).
Enzyme Catalysis and Regulation: A Model Enzyme Revisited
.
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Hatlem, Daniel; Leo, Jack Christopher; Saragliadis, Athanasios; Krengel, Ute & Linke, Dirk
(2017).
Bacterial adhesion: Characterization of trimeric autotransporter binding.
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Falao, Eirik & Krengel, Ute
(2023).
Studies of interaction between the Vibrio cholerae colonization factor GbpA and chitin using NMR spectroscopy and electron microscopy imaging.
Kjemisk Institutt, Universitetet i Oslo.
Show summary
he bacterium Vibrio cholerae is one of the most well-known contaminants of water and food sources across the world, and a cause of the disease cholera. It is particularly notorious for having outbreaks in underdeveloped parts of the world, or in areas affected by natural disasters or war, where access to clean drinking water is poor. Each year, cholera is estimated to cause 100 000 deaths. In addition to having the ability to colonize the human intestine, V. cholerae is known to colonize surfaces containing the polysaccharide chitin, found in the exoskeletons of several marine invertebrates. Colonization is believed to involve the adhesin N-acetyl glucosamine binding protein A (GbpA), which V. cholerae secretes into its aquatic environment. GbpA is a multifunctional protein with four domains, three of which have previously been structurally determined by x-ray crystallography. These are the N-terminal domain (D1), and the two middle domains (D2-3). D1 belongs to a class of enzymes called lytic polysaccharide monooxygenase (LPMO), which are known to cleave polysaccharides like chitin or cellulose. This domain is also known to interact with mucins, which are glycoproteins found in the human intestine. The functions of D2 and D3 are not known, but it has been hypothesized that they may play a role in anchoring V. cholerae to its substrate. The structure of the small C-terminal domain (D4) remains unknown, but it has been reported to also bind to chitinous surfaces. This thesis presents the results of interaction studies on GbpA with chitin, carried out by nuclear magnetic resonance (NMR) spectroscopy and electron microscope (EM) imaging. GbpA was produced using VmaxTM as an expression host. VmaxTM is a strain of Vibrio natriegens and a non-toxic relative of V. cholerae, possessing similar secretion pathways useful for GbpA expression and functionality. VmaxTM has previously been shown to express GbpA in high yields, which was confirmed in in this thesis. GbpA was also isotope-labelled with nitrogen-15 (15N) for NMR spectroscopy experiments. Although the NMR experiments yielded some inconclusive results, interactions between chitin and GbpA could be clearly observed in the EM-images obtained. Additionally, D4 was expressed alone with a glutathione S-transferase (GST) tag, using both Escherichia coli and VmaxTM as expression hosts. Although D4 was successfully isolated, extensive optimization of expression is required to acquire sufficient yields for v structure determination by NMR spectroscopy. An alternative method for isolating D4 was tested, where cleavage sites for TEV and HRV 3C proteases were inserted into the flexible linker separating D3 and D4, which would allow for GbpA to be expressed in high yields, from which D4 would be separated. Unfortunately, these cleavage sites could not be recognized by the proteases used, even though the same proteases could successfully cleave other proteins possessing the same cleavage sites. Predictions of the three-dimensional fold of D4 was made using AlphaFold and RoseTTAFold.
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Krengel, Ute & Montserrat Canals, Mateu
(2023).
Molecular warfare: A structural biology view on pathogen weapons — GbpA from Vibrio cholerae — and host defenses — Vg from the honey bee.
Universitetet i Oslo.
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Khatanbaatar, Tamjidmaa & Krengel, Ute
(2023).
Partners in crime: Structure-function studies on chorismate mutases and their associated enzymes.
Universitetet i Oslo.
ISSN 1501-7710.
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Hatlem, Daniel; Linke, Dirk & Krengel, Ute
(2022).
Probing the role of lipopolysaccharides in Gram-negative protein secretion and biogenesis.
Universitetet i Oslo.
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Sørensen, Henrik Vinther; Krengel, Ute; Lund, Reidar & Bjerregaard, Kaare
(2021).
Of Shellfish and Men: Applying X-ray and neutron techniques to investigate surface-active bacterial colonization factors.
Universitetet i Oslo, Det matematisk-naturvitenskapelige fakultet.
ISSN 1501-7710.
2021(2417).
Show summary
Bacteria responsible for diseases such as cholera and pneumonia form biofilms for surviving in the environment and inside their human hosts. The process involves enzymes that target sugars, providing the bacteria with nutrients. An in-depth understanding of the mechanisms of these enzymes can potentially lead to new vaccines, but also new means to convert complex sugars to clean energy sources. We used X-ray and neutron scattering methods to elucidate how these enzymes bind to chitin in the environment and how they degrade it. The work reveals new insight into the structures and mechanisms of the enzymes and how the bacteria use them to survive and thrive in the environment and inside humans.
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Pesci, Giacomo; Krengel, Ute; Johannesen, Hedda & Løset, Geir Åge
(2020).
Follow the Light - Tracking 14F7’s cellular journey with luminescence.
Universitetet i Oslo.
Show summary
Cancer is one of the leading causes of premature death in the world and its incidence and mortality are rapidly growing worldwide. Despite the biological heterogeneity of cancer, most patients are still treated with generic therapies, which often result in negative aftermath on the patients’ health. Adverse effects can be reduced by treatments targeting molecules which are exclusively expressed by cancer cells. One such molecule is the ganglioside N-glycolyl GM3 neuraminic acid (NeuGc GM3), which is present on the surface of several human cancer cell types and is not found in healthy tissue. This tumor-specific ganglioside is targeted with high specificity by the monoclonal antibody 14F7, which was produced in Cuba two decades ago. Unlike most anti-tumor antibodies, 14F7 has the outstanding ability to induce cancer cell death without recruiting any component of the immune system. To date, the cell killing mechanism of 14F7 is still poorly understood. In order to fully exploit its potential in cancer therapy, the fate of 14F7 after binding to the target has to be unraveled. The aim of this Master’s project was to gain insights into the internalization abilities of 14F7. After binding to the ganglioside, does the antibody travel inside the cell? Here, a 14F7 single-chain variable fragment (scFv) was covalently connected to a luciferase called NanoLuc. Due to its bright luminescence and pH-dependent activity, the NanoLuc has been identified as a suitable reporter for internalization studies. The vector coding for the fusion protein 14F7 scFv-NanoLuc was cloned through a PCR- based method, named Gibson assembly. A production protocol based on control of the glucose feed in Escherichia coli was established. The protein was effectively purified with immobilized metal affinity chromatography followed by size-exclusion chromatography. The binding affinity of the 14F7 scFv domain to NeuGc GM3 was confirmed by ELISA. The NanoLuc revealed a very high brightness and a robust pH- dependent activity. A recently developed assay to quantify the amount of a cell surface receptor, named Topanga assay, allowed quantification of NeuGc GM3 on several cell lines. Using the Topanga assay as a starting point, a novel assay to obtain insights into the internalization kinetics was developed. Interestingly, high amounts of NeuGc GM3 on the cell surface corresponded to faster internalization. This work paves the way for the application of scFv-C1-Nluc in bioluminescence microscopy in live cells.
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