|
|||||
|
|
Dynamics of Ligand Binding to a Rigid Glycosidase** |
|
| Authors: | Dr. Fredj Ben Bdira Dr. Christopher A. Waudby Dr. Alexander N. Volkov Dr. Sybrin P. Schröder Dr. Eiso AB Prof. Dr. Jeroen D. C. Codée Prof. Dr. Hermen S. Overkleeft Prof. Dr. Johannes M. F. G. Aerts Dr. Hugo van Ingen Prof. Dr. Marcellus Ubbink |
| Affiliation: | 1. Department of Macromolecular Biochemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC Leiden, The Netherlands;2. Institute of Structural and Molecular Biology, University College London and Birkbeck College, London, WC1E 6BT UK;3. VIB-VUB Center for Structural Biology, Pleinlaan 2, 1050 Brussels, Belgium Jean Jeener NMR Centre, VUB, Pleinlaan 2, 1050 Brussels, Belgium;4. Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC Leiden, The Netherlands;5. ZoBio BV, BioPartner 2 building, J.H. Oortweg 19, 2333 CH Leiden, The Netherlands;6. Department of Medical Biochemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC Leiden, The Netherlands |
| Abstract: | The single-domain GH11 glycosidase from Bacillus circulans (BCX) is involved in the degradation of hemicellulose, which is one of the most abundant renewable biomaterials in nature. We demonstrate that BCX in solution undergoes minimal structural changes during turnover. NMR spectroscopy results show that the rigid protein matrix provides a frame for fast substrate binding in multiple conformations, accompanied by slow conversion, which is attributed to an enzyme-induced substrate distortion. A model is proposed in which the rigid enzyme takes advantage of substrate flexibility to induce a conformation that facilitates the acyl formation step of the hydrolysis reaction. |
| Keywords: | dynamics glycosidases ligand binding NMR spectroscopy rigid fold |