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CCR7 signalling and ligand bias: A comprehensive characterisation using label-based and label-free methodologies KU Leuven

G protein-coupled receptors (GPCR) are intricate transmembrane proteins that relay signals from the extracellular to the intracellular environment using heterotrimeric G proteins. GPCRs mediate a vast array of physiological functions, but when deregulated, they also contribute to many pathological diseases such as diabetes, Alzheimer's disease, and cancer. This makes GPCRs an attractive therapeutic target. Indeed, approximately 35% of all Food ...

Radiosynthesis and preclinical evaluation of a 68Ga-labeled tetrahydroisoquinoline-based ligand for PET imaging of C-X-C chemokine receptor type 4 in an animal model of glioblastoma KU Leuven

Dominique Schols
BACKGROUND: This study aimed to develop a novel positron emission tomography (PET) tracer, [68Ga]Ga-TD-01, for CXCR4 imaging. To achieve this goal, the molecular scaffold of TIQ15 was tuned by conjugation with the DOTA chelator to make it suitable for 68Ga radiolabeling. METHODS: A bifunctional chelator was prepared by conjugating the amine group of TIQ15 with p-NCS-Bz-DOTA, yielding TD-01, with a high yield (68.92%). TD-01 was then radiolabeled ...

Development and application of NanoBRET-based technology: Unravelling signalling bias at the human chemokine receptors CXCR1 and CXCR2 KU Leuven

Katrijn Boon
G protein-coupled receptors (GPCRs) are cell surface receptors that translate extracellular stimuli into intracellular responses. They play a pivotal role in regulating developmental and physiological processes. Hence, dysregulated GPCR signalling contributes to various human pathologies, making GPCRs prime targets for FDA-approved drugs1-4. Ligand binding induces conformational changes in GPCRs, initiating both G protein-dependent signalling ...

Effects of Cure on the Ionic Conductivity and Relaxation Strength of a Reversible Polymer Network Studied by Dielectric Spectroscopy. Vrije Universiteit Brussel

Jessica Mangialetto, Niko Van den Brande, Michael Wubbenhorst
This study employs broadband dielectric spectroscopy (BDS) to investigate the cure kinetics and chemical equilibria of a Diels-Alder (DA) reversible network. Focusing on segmental dynamics, the α-relaxation and ionic conductivity are studied during heating and subsequent cooling right after isothermal precuring at 55 °C. During first heating, the glass transition temperature ( T g ) shows a systematic, stepwise upshift from -35 °C to 46 °C, ...

Insights in inulin binding and inulin oligosaccharide formation by novel multi domain endo-inulinases from Botrytis cinerea KU Leuven

Arnout Voet, Wim Van den Ende
World-wide, pathogenic fungi such as Botrytis cinerea cause tremendous yield losses in terms of food production and post-harvest food decay. Many fungi produce inulin-type oligosaccharides (IOSs) from inulin through endo-inulinases which typically show a two domain structure. B.cinerea lacks a two domain endo-inulinase but contains a three domain structure instead. Genome mining revealed three and four domain (d4) enzymes in the fungal kingdom. ...

Noncanonical and reversible cysteine ubiquitination prevents the overubiquitination of PEX5 at the peroxisomal membrane. KU Leuven

Marc Fransen
PEX5, the peroxisomal protein shuttling receptor, binds newly synthesized proteins in the cytosol and transports them to the organelle. During its stay at the peroxisomal protein translocon, PEX5 is monoubiquitinated at its cysteine 11 residue, a mandatory modification for its subsequent ATP-dependent extraction back into the cytosol. The reason why a cysteine and not a lysine residue is the ubiquitin acceptor is unknown. Using an established ...

Repurposing the mammalian RNA-binding protein Musashi-1 as an allosteric translation repressor in bacteria Vrije Universiteit Brussel

Roswitha Dolcemascolo, María Heras-Hernández, Lucas Goiriz, Roser Montagud-Martínez, Alejandro Requena-Menéndez, Raúl Ruiz, Anna Pérez-Ràfols, R Anahí Higuera-Rodríguez, Guillermo Pérez Ropero, Wim Vranken, et al.

The RNA recognition motif (RRM) is the most common RNA-binding protein domain identified in nature. However, RRM-containing proteins are only prevalent in eukaryotic phyla, in which they play central regulatory roles. Here, we engineered an orthogonal post-transcriptional control system of gene expression in the bacterium Escherichia coli with the mammalian RNA-binding protein Musashi-1, which is a stem cell marker with neurodevelopmental ...

Leucine Motifs Stabilize Residual Helical Structure in Disordered Proteins Vrije Universiteit Brussel

Uroš Zavrtanik, Tadej Medved, Samo Purič, Wim Vranken, Jurij Lah, San Hadži

Many examples are known of regions of intrinsically disordered proteins that fold into α-helices upon binding to their targets. These helical binding motifs (HBMs) can be partially helical also in the unbound state, and this so-called residual structure can affect binding affinity and kinetics. To investigate the underlying mechanisms governing the formation of residual helical structure, we assembled a dataset of experimental helix contents ...

PyRETIS 3 : conquering rare and slow events without boundaries Universiteit Gent

Wouter Vervust, Daniel T. Zhang, An Ghysels, Sander Roet, Titus S. van Erp, Enrico Riccardi
We present and discuss the advancements made in PyRETIS 3, the third instalment of our Python library for an efficient and user-friendly rare event simulation, focused to execute molecular simulations with replica exchange transition interface sampling (RETIS) and its variations. Apart from a general rewiring of the internal code towards a more modular structure, several recently developed sampling strategies have been implemented. These include ...

Modeling the effect of grain boundary diffusivity and trapping on hydrogen transport using a phase-field compatible formulation Universiteit Gent

Abdelrahman Hussein, Byungki Kim, Tom Depover, Kim Verbeken
Hydrogen grain boundary (GB) trapping is widely accepted as the main cause for hydrogen induced intergranular failure. Several studies were conducted to unveil the role of GBs on hydrogen transport; however, a clear understanding is yet to be attained. This is due to the limitations of the state-of-the-art experimental procedures for such highly kinetic processes. In this study, we aim at providing a deeper understanding of hydrogen-GB ...