Laboratory of Biomolecules Structure and Function ****************************************************************************************** * ****************************************************************************************** "Development and application of techniques for the structural description of clinical-rele biomacromolecules and the understanding of their structure-function modulation." ****************************************************************************************** * Offer ****************************************************************************************** We offer our expertise, as well as consultancy or collaboration within a diverse range of defined as protein biochemistry and biotechnology, protein structure solution or predictio spectrometry and biomacromolecule analyses, particularly in the fields of: - Recombinant protein expression and purification (both in bacterial and in mammalian cell systems) - Protein characterization and binding assessment (by various biochemical and biophysical sedimentation analysis in analytical ultracentrifuge, surface plasmon resonance, UV-VIS & - Protein structure determination (by protein X-ray crystallography or by advanced techniq spectrometry) - Multi-protein dynamics mapping - Molecular dynamic and protein structure modelling ****************************************************************************************** * Know-how & Technologies ****************************************************************************************** Structure and function relationship of biomacromolecules: target biomolecule identificatio characterization, biotechnologies dealing with recombinant proteins as therapeutic agents. - Recombinant protein expression and purification - Structural biochemistry and biophysical characterization of proteins - Structure of ligand-protein and multi-protein complexes - Photo-initiated and chemical cross-linking, H/D exchange - Mass spectrometry of biochemically interesting macromolecules - Molecular modelling of biomolecules ****************************************************************************************** * Main Capabilities ****************************************************************************************** Consultancy or collaboration within the fields of recombinant protein expression and purif in bacterial and in mammalian cell line expression systems) and protein characterization b biochemical and biophysical techniques, e.g. sedimentation analysis in analytical ultracentrifuge, surface plasmon resonance, UV-VIS & structure solution by protein X-ray crystallography or by advanced techniques of mass spec *========================================================================================= * Content of Research *========================================================================================= The structural and functional characterization of proteins participating in cancer recogni xenobiotic biotrans-formation, in the regulation of cell processes, in biocatalysis and im in their native conditions. Research of protein structure, protein-protein and protein-lig interaction with the use of chemical cross-linking reagents, photo-initiated cross-linking photoaffinity labeling protein, determination of disulfide bridges, glycosylation and othe translational modifications of proteins. Protein identification, structural determination advanced techniques of mass spectrometry.Homology modeling, in silico ligand docking, mole and interaction energy calculations. *========================================================================================= * Research Equipment *========================================================================================= All equipment is operated by responsible persons: M. Šulc, M. Martínková, V. Martínek, P. Vaněk, D. Kavan, P. Pompach, P. Jeřábek, T. Ječmen, R. Ptáčková, J. Bláha. - Analytical ultracentrifuge ProteomeLab XL-I - Surface plasmon resonance workstation PLASMON-4 - HPLC/UHPLC and FPLC systems - UV-VIS spectrophotometers, CD spectrophotometer - Protein sequencer Procise 491 - Two photolysers emitting UV-light with maximum around 254 nm (100W) and 360 nm (500W) - Bruker Daltonics 15T-Solarix XR FT-ICR mass spectrometer associated with Agilent Technol system - Bruker qTOF maXis Plus ****************************************************************************************** * Partners and Collaborations ****************************************************************************************** *========================================================================================= * Academic Partners *========================================================================================= University of Texas, Houston, USA | Technische Universität Berlin, Germany | Institute for Science, Okazaki, Japan | National Institute of Molecular Studies, Okazaki, Japan | Loyola Chicago, USA | Georgetown University, Georgetown, USA *========================================================================================= * Public and Private Sector *========================================================================================= Zentiva, a.s. | Apronex, s.r.o. *========================================================================================= * Main Projects *========================================================================================= - New ligands for old receptors of human natural killer cells: structure, assembly within synapse and potential for therapy (Czech Science Foundation, 2018–2021) - Utilization of integrative molecular biophysical approaches for the functional studies o killer cell receptors and formation of their complexes with tumour ligands (INTER-COST MEY - Molecular mechanisms of intraprotein/interdomain signal transduction in model heme senso Science Foundation, 2015–2017) - Harnessing soluble forms of NK cell receptors and their ligands for the generation of no immunotherapeutics (Czech Science Foundation, 2015–2017) - Innovative Technologies for the Identification and Optimization of the New Generation of Drugs (Charles University, UNCE 204025/2012, 2012–2017) - Mammalian microsomal cytochrome P450 interaction with redox partners – topology and stru relationships (Czech Science Foundation, 2012–2015) - Innovation voucher (with Zentiva, a.s., Prague city, 2013–2014) - Molecular diagnostics of bacterial pathogens (with Apronex, s.r.o., Ministry of Industry 2013) ****************************************************************************************** * Achievements ****************************************************************************************** *========================================================================================= * Patents *========================================================================================= - Czech patent “Soluble form of mouse NK cell receptor NKR-P1A and means of its recombinan preparation” (PV 2010-132) - Czech utility model “Active form of alfa-N-acetyl-galactosaminidase from filamentous fun niger” (2012-26061) *========================================================================================= * Recent Publications *========================================================================================= - Stranava M et al.: Coordination and redox state–dependent structural changes of the heme sensor AfGcHK associated with intraprotein signal transduction. J Biol Chem. 292, 20921–93 - Kadek A.: Interdomain electron transfer in cellobiose dehydrogenase is governed by surfa electrostatics. Biochim Biophys Acta. 1861, 157–167 (2016) - Vališ K.: Reprogramming of leukemic cell metabolism through the naphthoquinonic compound Oncotarget. 8, 103137–153 (2017) - Bláha J.: High-level expression and purification of soluble form of human natural killer NKR-P1 in HEK293S GnTIcells. Protein Expr Purif. 140, 36–43 (2017) - Škerlová J.: Crystal structure of native ?-N-acetylhexosaminidase isolated from Aspergil light onto its substrate specificity, high stability, and regulation by propeptide. FEBS J (2018) - Ječmen T.: Photo-initiated crosslinking extends mapping of the protein-protein interface embedded portions of cytochromes P450 2B4 and b5. Methods. 89, 128–37 (2015) *========================================================================================= * Are you interested in this expertise? *========================================================================================= Please contact CPPT UK Web: www.cppt.cuni.cz/ [ URL "https://cppt.cuni.cz/"] Mail: transfer@cuni.cz Phone: +420 224 491 255 *========================================================================================= * Experts and their department *========================================================================================= Assoc. Prof. Miroslav Šulc, Ph.D. Department of Biochemistry