The Institute of Interdisciplinary Research (IRIBHM) is part of the ULB Medical School. About 140 researchers and technicians are working in the Institute over a range of subjects essentially focused on cell signalling, using cell and molecular biology approaches. About one third of the Institute is working on GPCRs and is concerned by the present network proposal. This group has pioneered homology cloning of G protein-coupled receptors, and a large number of original receptors have been isolated and characterised functionally using this approach (including thyrotropin, adenosine A1 and A2a, purinergic P2Y4, P2Y11 and P2Y13, chemokine CCR5, NPFFR2, GPR43, among others). The group also purified a number of natural agonists of orphan receptors, including a truncated HCC-1 variant for CCR5, kisspeptins for GPR54, chemerin for ChemR23 and the F2L peptide for FPRL2. The group identified genetic alterations of GPCRs causing human diseases (thyrotropin receptor in hyperfunctional adenoma, FSH receptor in ovarian hyperstimulation syndrome), or protecting individuals against HIV infection (CCR5), and developed over 20 transgenic (addition or knock out) mouse models so far. In the frame of this past activity, the group has gained a wide experience in all aspects of GPCR characterisation. The heavy equipment available in the Institute includes a mass spectrometer, diverse HPLC systems, 2D-DIGE electrophoresis and analysis system, FACS, fluorescence microscopes, confocal microscopes, DNA sequencers, beta and gamma counters, a range of plate luminometers and fluorimeters, robotic workstations, DNA arrayer, microarray scanner, videotracking and other equipment for behavioural analysis, microinjection equipment for cell cultures and mouse embryos, and a SPF animal facility.
The 'Laboratory of Molecular Cell Biology' is part of the Department of Biology and is also the 'Department of Molecular Microbiology' of the VIB. It counts about 60 researchers of which about 20 are involved in nutrient-sensing research. It has 25 years of experience in the elucidation of the molecular genetics and cellular physiology of nutrient sensing and signalling in yeast. It developed the best understood multi-nutrient sensing system in eukaryotic cell biology and discovered the first nutrient-sensing GPCR and the first examples of nutrient transceptors. It has experience in expressing and characterizing mammalian genes in yeast and in development of specific screens with yeast mutants to isolate functional mammalian substitutes of yeast genes. SCAM (Substituted Cysteine Accessibility Method) analysis of GPCRs and transporter-receptors. Determination of cAMP, metabolites, enzyme activities, cellular transport measurements and various parameters of yeast physiology. Use of radioactive tracers. GC and HPLC analysis. FACS. Confocal microscopy for protein localization in living cells. Purification of proteins. All modern methods of recombinant DNA technology, gene expression analysis using microarrays and QPCR, immunological methods, immunofluorescence, two-hybrid and split-ubiquitin screening, methods of yeast transformation, mutant and suppressor selection and identification, gene deletion and overexpression in yeast, site-directed mutagenesis and polygenic analysis of complex traits using pooled segregant whole genome sequencing.
The group of Jozef Vanden Broeck is one of the leading groups in the world in insect endocrinology and neurobiology. They pioneered the discovery of novel peptides, peptide precursors and G protein-coupled receptors. Their overall objective is to study the molecular signalling components involved in important developmental-physiological processes from an evolutionary perspective. Relevant technical expertise: molecular biology, cellular expression technology, receptor and signal transduction studies, reporter assays, fluorometry, luminometry, chromatography (HPLC) and sequencing of peptides, proteomics and mass spectrometry, electrophoretic methods, microfluidics, DNA sequencing, nucleic acid hybridizations, immunological localization and detection methods, peptidase and peptidase inhibitor assays, microarrays, realtime PCR, bio-informatics applications, insect cultures, behavioural and physiological assays, RNA interference, Drosophila molecular genetics.http://bio.kuleuven.be/mcb/
The Laboratory of Molecular Immunology belongs to the Department of Microbiology and Immunology of the Faculty of Medicine. Our group has a long tradition in cytokine research in relation to inflammation, infection and cancer and is involved in G protein-coupled receptors through on the discovery of inflammatory mediators, such as cytokines (interleukin-1 and -6) and chemokines (CXCL6, CXCL8, CCL2, CCL7, CCL8). The laboratory has the skills to produce, isolate and identify novel mediators from in vitro stimulated cell cultures or body fluids from patients with infection, autoimmune diseases or cancer. These include large scale isolation and/or culture of various cell types in vitro and chromatographical purification of secreted proteins and posttranslational modifications thereof to homogeneity and their identification through amino acid sequence analysis and mass spectrometry. This can also be implemented on smaller scale, for instance on patient’ samples. Specific immunotests are developed to measure cytokines and chemokines in patients and to study their gene regulation in vitro. New chemokine isoforms are chemically synthesized on large scale to perform in vivo experiments using models of inflammation or cancer. Chemokines are evaluated for their capacity to influence angiogenesis, to chemoattract various leukocyte subtypes (freshly isolated from blood) and to activate GPCR in different signal transduction assays. To perform these experiments we have up-to-date instrumentation available, including: HPLC systems and a 2D nanoHPLC with online mass spectrometry, amino acid sequencing and peptide synthesis equipment, flow cytometry, quantitative RT-PCR, fully equipped fluoresence microscopy for live cell imaging and two-photon confocal microscopy units, and an SPF animal facility.http://www.kuleuven.be/onderzoek/onderzoeksdatabank/onderzoeksgroep/50000703.htm
The Jan Steyaert lab (http://steyaertlab.structuralbiology.be) is part of Structural Biology Brussels at the VUB and of the Department of Structural Biology of the VIB. Jan Steyaert was trained as a structural enzymologist but changed gears to focus on the use of nanobodies to facilitate the crystallization of challenging targets. His lab identifies Nbs that stabilize unique conformations of membrane proteins and transient protein-protein interactions to study their structure, function and dynamics. Within our Nanobody platform, we immunize llamas with purified protein, with cells expressing the target or by genetic vaccination to induce an in vivo matured humoral respons against native conformations of the protein of interest. Using phage display, we select diverse panels of in vivo matured Nbs that facilitate the crystallization of the protein of interest. We identify Nbs that selectively stabilize native conformations of these targets. Within our structural biology platform, we purify stable Nb-protein complexes that are amenable to crystallisation. Our lab is fully equipped with liquid handling and nanodispensing crystallization robots for high throughput Nb co-crystallization screens and condition optimization. We screen crystals and collect data on our in house X-ray generator to solve the structures of the stabilized Nb-protein complexes. Our lab is also outfitted with 600 and 800 MHz NMR spectrometers, equipped with a salt-tolerence cryoprobe for biomolecular applications.http://www.steyaertlab.be
The Laboratory of Immunology belongs to the Department of Biomedical Science and Biotechnology of the Faculty of Medicine. Our group has a long lasting tradition in the characterization of new chemotactic factors and in the study of the role of chemokines in inflammation and autoimmune diseases (e.g. Vermi et al. J Exp Med, 2005; Albanesi et al. J Exp Med, 2009; Parolini et al Blood, 2007). In the past we have contributed to the understanding of the role of chemokines in dendritic cell trafficking publishing the very first report about chemokine regulation of DC migration (Sozzani et al J. Immunol., 1995), formulating the paradigm of chemokine receptor switch during DC maturation (Sozzani et al. J Immunol, 1998), participating to the biological characterization of new chemokines (e.g. CCL20 and CCL22) and chemokine receptors (i.e. CCR6 and ChemR23) (Godiska et al., J Exp Med 1997; Power et al., J Exp Med 1997; Wittamer et al., J Exp Med 2003) and providing the first description of the selective usage of chemokine receptors by blood plasmacytoid DC (Penna et al., J Immunol, 2001). A particular interest was also directed to the study of second messengers involved in DC migration. Phospholipase A2 and phospholipase D were studied for their role in chemokine signal transduction (Locati et al. J Biol Chem, 1994; Locati et al., J Biol Chem, 1996; Locati et al. Biochem J, 2001). The non-redundant role of PIP3K in phagocyte migration, including DC, was also reported (Hirsh et al., Science, 2000; Del Prete et al., EMBO J, 2004). This laboratory is fully equipped for tissue culture, ELISA, RealTime PCR, FACS analysis (Partec) and fluorescent time-lapse microscope (Zeiss).