Spectroscopy of biomolecules
Spectroscopy of biomolecules
Academic year 2017/2018
- Course ID
- Teaching staff
- Simonetta Geninatti Crich
Prof. Gianmario Martra
- Modular course
- Degree course
- [0101M21] Molecular Biotechnology
- 1st year
- Course disciplinary sector (SSD)
- CHIM/02 - chimica fisica
CHIM/03 - chimica generale e inorganica
- Formal authority
- Type of examination
- Basic knowledge in Inorganic Chemistry, Physical Chemistry, Biochemistry
- Propedeutic for
- All courses and practical activities dealing also with methods based on molecular spectroscopies
Sommario del corso
The objectives proposed to students in this course are based on the following achievements:
1. knowledge of the chemical, physical and physical-chemical principles at the basis of the generation of signals exploited in molecular spectroscopies of biomolecules, namely NMR spectroscopy and electron spectroscopies (both in absorption and emission)
2. good capability in critical understanding scientific texts
3. good capability in designing investigation of different samples by molecular spectroscopies, on the basis on their constitutive features, possible constrains related to limited amounts, types of outputs requested (identification; quantification)
4. good capability in the complementary use of the molecular spectroscopic methods indicated above
Results of learning outcomes
- Knowledge of main sample handling methods for NMR and electronic spectroscopic (both in absorption and emission) measurements
- Capability to design proper spectroscopic measurements for the investigation of biomolecular samples, taking into account the peculiar features of these samples (e.g, complexity of the matrix), and the specific target(s) to be pursued (identification/quantification)
- Capability to analyse data resulting from the methods indicated above.
The course will be delivered through lessons in the lecture room (90%) and in labs (10%), for focus on some methodological aspects.
A half of the coiurse will be devoted to NMR (3 CFU, 24 h) and a half to electronic (absorption and emission) spectroscopy (3 CFU, 24 h)
Learning assessment methods
The exam is devoted to the assessment of the knowledge, and related understanding, of the program attained by the students. In addition, also knowledge and understanding of basic knowledge in Chemistry and Biochemistry which should be necessary consider will be evaluated, as well as the use of a proper scientific/technical language.
The examination is carried out in oral form. The final mark is expressed in thirtieths.
Absorption electronic spectroscopy:
- origin of the energy transfer from the electromagnetic radiation to molecular electronic states
- types of electronic levels and electronic transitions
- UV circular dichroism
- study of the protein structure by absorption electronic spectroscopy (principles, experimental methods)
- study of the structure of nucleic acids by absorption electronic spectroscopy (principles, experimental methods)
Photoluminescence electronic spectroscopy:
- radiative and radiation less decays from excited electronic states
- fluorescence and phosphorescence
- radiative and fluorescence lifetimes
- collisional quenching
- energy transfer through the space
study of the protein structure by photoluminescence spectroscopy (principles, experimental methods)
- study of the structure of nucleic acids by photoluminescence spectroscopy (principles, experimental methods)
Nuclear Magnetic Resonance (NMR) Spectroscopy applications in the determination of structure, dynamics, and interactions of biological macromolecules:
- Introduction to the NMR spectral parameters used in structural biology, namely the chemical shift, the J-coupling, nuclear Overhauser effects, and residual dipolar couplings.
- Resonance assignment, and NMR spectral parameters are their convertion into angle and distances between atoms
in a macromolecules.
-Relaxation phenomena and mapping the molecular interactions, information on the binding interface as well as the determination of kinetic and thermodynamic constants..
-NMR studies on Metalloproteins (proteins containing at least one metal ion) and paramagnetism based drug discovery.
- Nucleic acids studied by NMR and their interaction with metallo drugs.
- In cell/In tissue NMR spectroscopy.
Suggested readings and bibliography
Reference material is available at the course website.
Suggested textbooks (all available from the teachers):
- H. Friebolin “Basic one- and two-dimensional NMR spectroscopy”, VCH, 1993
- Ivano Bertini, Kathleen S. McGreevy, Giacomo Parigi “NMR of biomolecules” Wiley 2012.
- DW. Claridge “High-resolution NMR techniques in Organic Chemistry”, Pergamon 1999 (Elsevier Science)
N.J. Turro, Modern Molecular Photochemistry, University Science Books, 1991
C.N.R. Rao, Ultra-Violet and Visible Spectroscopy, Butterworths, third edition, 1966
J.R. Lakowitz, Principles of fluorescence spectroscopy, Plenum, 2000 I.D. Campbell