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Lecturer(s)
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Šebela Marek, prof. Mgr. Dr.
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Course content
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Protein analysis, determination of characteristic parameters (Mr, pI, subunit composition). Determination of protein concentration. Computer programs for prediction of protein properties using data retrieved from databases, ExPASy server. Separation of protein mixtures -chromatographic and elecrophoretic methods. Detection techniques for proteins. Spectroscopic analysis of proteins. Protein crystallography, protein crystallization, X-ray diffraction analysis, phase problem and its solving. NMR spectroscopy of proteins, multidimensional NMR techniques, assigning of spectra. Mass spectrometry of proteins, determination of molecular mass, analysis of post-translational modification (PTM), utilizing mass spectrometry for study of 3-D protein structure (ligands, bifunctional reagents for crosslinking). Protein sequencing, Edman chemistry, mass spectrometry-assisted sequencing (fingerprinting, de novo sequencing). Computer modeling of protein structure, protein structure database. Study of proteins in non-aqueous environment, encapsulation, microemulsion, preparation of lipid-protein aggregates (ELA). Techniques of protein immobilization. Enzyme reactors, biosensors. Chemical and enzymatic modification of proteins. Modification for increased thermostability, resistance against proteolysis, modulations of activity and specificity of enzymes. Reagents for specific modification of amino acids. Low-molecular-weight modification reagents, bifunctional reagents for crosslinking. PEGylation of proteins, attachment of hydrophilic polymers. Artificial glycosylation of proteins. Analysis of modification extent.
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Learning activities and teaching methods
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Dialogic Lecture (Discussion, Dialog, Brainstorming), Work with Text (with Book, Textbook)
- Preparation for the Exam
- 50 hours per semester
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Learning outcomes
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Students will acquaint themselves with basic and advanced methods of studying and analyzing proteins including chemical modifications.
Explain main aspects of the chemistry of proteins, predict chemical properties of proteins.
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Prerequisites
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The candidate is expected to conduct scientific discussion in broader context of the whole discipline.
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Assessment methods and criteria
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Oral exam
the candidate is expected to conduct scientific discussion in broader context of the whole discipline.
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Recommended literature
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Buxbaum E. (2011). Biophysical Chemistry of Proteins. An Introduction to Laboratory Methods. Springer.
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Howard G.C., Bron,W.E. (2002). Modern Protein Chemistry. Practical Aspects. CRC Press, Boca Raton, FL.
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