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Lecturer(s)
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Kutá Smatanová Ivana, prof. Mgr. Ph.D.
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Maloy Řezáčová Pavlína, doc. RNDr. Ph.D.
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Kaščáková Barbora, Mgr. Ph.D.
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Havlíčková Petra, RNDr.
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Course content
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Content of lectures: Macromolecular substances - basic terms Crystallization - principles and methods of crystallization Properties of solutions and phase diagrams Automation in crystallization of proteins Nucleation and crystal growth, growth kinetics Membrane proteins crystallization Standard crystallization methods Advanced crystallization methods Alternative crystallization methods Mounting and freezing of crystals Interpretation of crystallization experiments results Publication of crystallization results Content of exercises: Methods of protein crystallization, crystallization experiments with model proteins (eg crystallization of lysozyme, proteinase K, etc.), familiarization with the work of crystallization robots, preparation of protein crystals for measurement on a diffractometer, introduction to the structure solution of protein using PX software.
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Work with text (with textbook, with book), Demonstration, Laboratory, Activating (simulations, games, drama)
- Class attendance
- 65 hours per semester
- Preparation for credit
- 10 hours per semester
- Preparation for exam
- 10 hours per semester
- Preparation for classes
- 10 hours per semester
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Learning outcomes
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The X-ray crystallography is becoming one of the most important techniques to study proteins, nucleic acids and their complexes as well as viruses and supramolecular complexes at an atomic resolution. Crystallization of biomacromolecules is an empirical process based on finding the individual conditions and parameters for growing crystals and their optimization. The limited condition of crystallization is growing crystals suitable for X-ray diffraction measurement. The course is aimed at familiarization of students with standard and advanced crystallization methods and techniques. During the lab exercises students will grow crystals of model or own proteins using of standard and advanced crystallization techniques.
Graduates will became familiar with methods and techniques of crystallisation of important biomolecular compounds like proteins and nucleic acids. They will be able to find by own individual conditions and parameters for growing crystals and their optimalization using standard, advanced and alternative crystallisation techniques, moreover they will know how to test character of crystals and prepare crystals for diffraction measurements on the X-ray sources (diffractometers and synchrotrons) for a final determination of macromolecular structure.
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Prerequisites
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Completion of a general and organic chemistry and biochemistry courses and laboratory work.
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Assessment methods and criteria
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Student performance assessment, Combined exam, Development of laboratory protocols
Requirement for successful completion of the course is the active participation of the student in practical exercises. The final exam will take the form of a written test and a subsequent oral interview with a final mark in the form of a mark.
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Recommended literature
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A.Ducruix; R. Giege. Crystallization of nucleic acids and proteins : a practical approach 2Ed. Oxford University Press, 2000. ISBN 9780199636785.
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Daniel M. Bollag, Michael D. Rozycki, Stuart J. Edelstein. Protein Methods, 2nd Edition. Wiley, 1996. ISBN 978-0-471-11837-4.
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Jan Drenth. Principles of Protein X-Ray Crystallography 2Ed. Springer Verlag Publishing, 2002. ISBN 978-0-387-33746-3.
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T.M. Bergfors. Protein Crystallization. Techniques, Strategies, and Tips. International University Line, 1999. ISBN 0963681753.
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