1. Introduction to Molecular Spectroscopy. Interaction of radiation with matter. Types of radiation and spectra. General characteristics.
2. Nuclear Magnetic Resonance Spectroscopy. Introduction to physical bases of NMR. Spectral parameters and their information content. Examples in biological systems. Chemical displacement. Coupling constant. Magnetic excitation and relaxation. One-and two-pulse experiments. Chemical exchange effects. Magnetization transfer. Two-dimensional NMR. Applications in structure elucidation of small molecules, peptides, proteins, nucleic acids and biological membranes. Metabolic and clinic studies by NMR. Introduction to medical magnetic resonance imaging (MRI). Contrast agents.
3. Spectroscopy of Electron Paramagnetic Resonance. Physical bases and instrumentation. Spectral parameters and their information content. g-value, relaxation and hyper-thin structure of spectra. Applications in Biology and Medicine. Spin traps. Spin probes in Biophysics. Transition metal RPE in metalloproteins.
4. Electron absorption spectroscopy and circular dichroism. Chromophors in ultraviolet-visible. Instrumental aspects. Spectra parameters. Rules of selection and intensities of electronic transitions.
Biopolymer spectra – proteins, nucleic acids. Hypochromism. Optic activity of electronic transition bands – optical rotation and circular dichroism. Cotton effect. Secondary structure of proteins from circular dichroism.
5. Luminescence Spectroscopy. Fluorescence and phosphorescence. Inter-systems conversion. Life times and transition between states. Nonradiative deactivation processes. Stokes deviation. Environmental effects. Fluorescence quantum efficiency. Factors that affect fluorescence intensity. Polarity probes. Biophysical applications. Dynamic Quenching. Controlled diffusion processes. Applications in Biophysics.
6. Infrared spectroscopy. Experimental parameters. Hooke’s Law and harmonic oscillator. Vibrational energy levels. Zero-point energy. Non-harmonic vibrations. Vibrational transitions. Normal modes of vibration. Polyatomic molecules. Groups characteristic frequencies. Digital printing region. Examples and applications.
7. Light scattering techniques. Light scattering by matter. Rayleigh elastic scattering. Scattering by a network of particles. Light scattering and diffraction by crystals – X-ray, neutrons – atomic factors. Bragg equation. Diffractograms and maps of electron density. Electron diffraction and electron microscopy. Light scattering in solution. Inelastic scattering – dynamic and Raman. Vibrational transitions. Resonance Raman spectra.
8. Mass spectrometry. Introduction to the physical bases of mass spectrometry. Biological applications. Protein structure and interaction. Proteomics and metabolomics.
Bibliography of reference
CAMPBELL, I. D.; DWEK, R. A (1984). Biological Spectroscopy, Benjamin. California: Menlo Park.
HOLDE, K.E. Van (1985). Physical Biochemistry. N. Jersey: Prentice Hall.
CANTOR C. R.; SCHIMMEL, P. R. (1980). Biophysical Chemistry, Part II: Techniques for the Study of Biological Structure and Function. New York: Freeman.
DRAGO, R.S. (1992). Physical Methods for Chemists. Ft. Worth: Saunders.
WÜTHRICH, K. (1986). NMR of Proteins and Nucleic Acids. Willey and Sons, Inc.
KEELER, J. (2005). Understanding NMR Spectroscopy. Willey and Sons, Inc.
EVANS, J.N.S (1995). Biomolecular NMR Spectroscopy. Oxford University Press.
RULE, G.S.; HITCHENS, T.K. (2005). Fundamentals of Protein NMR Spectroscopy in Structural Biology. Springer.
KRISHNA, N.R.; BERLINER, L.J. (2003). Protein NMR for the Millenium. Springer.
I. A., Kaltashov; S. J. Eyles (2005). Mass Spectrometry in Biophysics: Conformation and Dynamics of Biomolecules. Willey and Sons, Inc.