- Laboratory of Enzymology and Protein Folding
Centre d’Ingénierie des Protéines (CIP)
Bât.B6c Quartier Agora
allée du six Août 11
4000 Liège 1
- +32 4 3663419
- +32 4 3663364
- Molecular Biophysics
- Protein Chemistry
- Protein Folding
Understanding the basic aspects of Protein Folding is crucial in describing many cellular processes, ranging from transcription to molecular motors and diseases associated with misfolded proteins. Rational modification and de novo conception of novel proteins with therapeutic or biological applications, and prediction of their three-dimensional structures from their amino acid sequence, require both a detailed description of the energetics of folding and knowledge of the driving forces and pathways that lead to the native state. Much of the progress in understanding the way in which proteins fold has been marked by intensive studies on model proteins. Proteins such as ribonuclease, lysozyme and bovine pancreatic trypsin inhibitor were chosen for their availability, for particular points of structural interest, and for their solubility and ready reversibility of unfolding in the test tube.
Although the events in the folding of globular proteins appear to be diverse and complex, a wealth of information has been gathered from over fifty years of research. Nevertheless, some details of the folding mechanism are still unclear and, in particular, prediction of the sequence of acquisition of secondary and tertiary structural elements of multi-domain proteins (> 100 amino acids) remains a difficult task.
With the goal of contributing to a better description of the protein folding problem, we have selected model proteins with essentially different folds. These include active-site serine β-lactamases (α + β structure ; Mr ~29000), Zn(II) metallo-β-lactamases (α + β structure ; Mr ~25000), lysozymes (α + β structure; Mr ~14000-18000), single domain antibody fragments from camelid heavy chain antibodies (VHH; all-β structure ; Mr ~15000) and Erwinia chrysanthemi pectine methylesterase (mainly β structure, organized into a right-handed parallel β-helix ; Mr = 36899). Folding of these proteins is analyzed by using a range of complementary spectroscopic probes. Thus, optical methods (i.e. fluorescence and circular dichroism) in combination with rapid-mixing techniques provide a first description of the folding mechanism. In the case of enzymes, this can be advantageously completed by measuring the regain of catalytic activity. Furthermore, we use pulse-labelling hydrogen/deuterium exchange experiments, in combination with 2D-NMR (collaboration with Christina Redfield, University of Oxford and Christian Damblon, University of Liège) and/or mass spectrometry (collaboration with Edwin De Pauw, University of Liège) measurements, to monitor the time-course of formation and stabilization of secondary structure elements.
Finally, in a collaborative effort with Cécile Van de Weerdt (University of Liège) and Maximiliano Figueroa (University of Conception), we work on the design of proteins with an (β/α)8, known as Octarellins.
- Julie Vandenameele, Postdoctoral fellow
- Kishore Babu Bobbili, Postdoctoral fellow
- Cristina Martina, PhD student
- Ruth Kellner, Postdoctoral fellow
- Matagne, A., Misselyn-Bauduin, A.M., Joris, B., Erpicum, T., Granier, B. and Frère, J.M. (1990) The diversity of the catalytic properties of class A β-lactamases. Biochem. J. 265, 131-146.
- Matagne, A., Radford, S.E. and Dobson, C.M. (1997) Fast and slow tracks in lysozyme folding: insight into the role of domains in the folding process. J. Mol. Biol. 267, 1068-1074.
- Dumoulin, M., Conrath, K., Van Meirhaeghe, A., Meersman, F., Heremans, K., Frenken, L.J.G., Muyldermans, S., Wyns, L., and Matagne, A. (2002) Single-domain Antibody Fragments with High Conformational Stability. Protein Sci. 11, 500-515.
- Dumoulin, M., Last, A. M., Desmyter, A., Decanniere, K., Canet, D., Larsson, G., Spencer, A., Archer, D.B., Sasse, J., Muyldermans, S., Wyns, L., Redfield, C., Matagne, A., Robinson, C.V. and Dobson, C.M. (2003) A camelid antibody fragment inhibits the formation of amyloid fibrils by human lysozyme. Nature 424, 783-788.
- Di Paolo, A., Balbeur, D., De Pauw, E., Redfield, C., Matagne, A. (2010) Rapid Collapse into a Molten Globule Is Followed by Simple Two-State Kinetics in the Folding of Lysozyme from Bacteriophage λ. Biochemistry 49, 8646–8657.