Laboratory of elementary biophysics S 1100-3BP21

Obligatory experiments
1. UV/VIS absorption spectroscopy
2. Fluorescence microscopy (FLIM)
3. UV/VIS emission spectroscopy
4. Kinetics and inhibition of enzymatic reactions
5. NMR spectroscopy
6. Experiments that make use of microplate reader

Experiments to choose (not all available an each academic year)
1. Ionic forms, spectrophotometric determination of pK and pK*
2. Spectroscopy of circular dichroism (CD)
3. Kinetic studies using the stopped-flow method
4. Proteins unfolding
5. Fluorescence titrations
6. Isotope methods
7. Chromatographic methods
8. Life-times of excited states
9. Mass spectrometry (MS)
10. Spectroscopic studies of tautomeric forms of nucleic acid components.
11. Infrared spectroscopy
12. Isothermal titration calorimetry (ITC)
13. Analytical ultracentrifugation
14. Crystallization of proteins

Mode

Classroom

Prerequisites (description)

Laboratory is available for students who passed the following exams Physical chemistry, Laboratory of chemistry, Laboratory of molecular biology, Physics and mathematics, Analysis of physical measurements, Physics of atoms, molecules and biological macromolecules, Structure and function of biological macromolecules, Molecular spectroscopy, Methods of molecular biophysics. The main goal of Laboratory of elementary biophysics is to introduce students of specialization Molecular biophysics to modern experimental methods used in molecular biophysics. Theoretical bases of these methods were described in the frame of two previous courses: “Molecular spectroscopy” and “Methods of molecular biophysics”. The laboratory consists of short, 6-hours, one-day experiments that are conducted together by two students. In the case of equipment such as spectrometer NMR by two such groups (four students). The course of all experiments is well defined to use in the optimal way the short time devoted to each experiment.

Learning outcomes

Knowledge:
1. Basis of molecular spectroscopy
2. Structure and spectroscopic properties of nucleosides, nucleotides, nucleic acids, aminoacids and proteins
3. Spectroscopic methods used in biophysical studies
4. Methods to analyze protein-ligand interactions
5. Kinetics and inhibition of enzymatic reactions
6. Basis of analytical ultracentrifugation
7. Methods for crystallization of proteins

Skills:
1. To conduct simple experiments in molecular biophysics
2. To work in team
3. To use basic equipment useful in biophysical studies
4. To analyze and present the results of experiments

Attitudes:
1. Understanding what means ethics in science
2. Responsibility for results of experiments
3. Proper behaviour while working with chemical and biological samples and equipment use in biophysical studies

Assessment criteria

Credits for the Laboratory are given based on:
1. positive notes for entrance colloquia
2. performance during classes
3. positive notes for laboratory reports that were prepared individually by each student

Maria Agnieszka Bzowska, last modification November 25, 2011

Bibliography

1. G. Ślósarek, „Molecular biophysics. Phenomena. Instruments. Modeling”, PWN, Warszawa
2. Z. Kęcki, „Basis of molecular spectroscopy”, PWN, Warszawa
3. P.W. Atkins, „Physical chemistry”, PWN, Warszawa
4. Praca zbiorowa, „Spectroscopic methods and their application to identification of organic molecules ", WNT, Warszawa
5. A. Kawski „Photoluminescence of solutions”. PWN, Warszawa
6. R.A. Copeland, „Enzymes” Wiley-VCH, New York
7. A. Fersht, „Structure and mechanism in protein science” W.H. Freeman and Company, New York
8. G.U. Nenhaus „Protein-ligand interactions. Methods and applications” Human Press, Totowa, New Jersey.
9. C.A. Parker „Photoluminescence of solutions” Elsevier, Amsterdam
10. J.R. Lakowicz, „Principles of Fluorescence Spectroscopy”, Kluwer Academic/Plenum Press Publisher, New York, Boston, Dordrecht, London, Moscow