Structure elucidation of peptides and proteins by NMR
Kém/251
Structure elucidation of peptides and proteins by NMR
The aim of the lecture is to introduce the theory and practice of NMR-spectroscopy of organic- and biomolecules. The principles of how to evaluate complex spectra and do structure elucidation are provided. Theoretical basis of bio-NMR spectroscopy are given to start with, such as: the vector model, the product operator formalism, relaxation, nuclear Overhauser effect, polarisation transfer, scalar and dipolar coupling, population and coherence transfer, chemical shift etc..
NMR signal assignment and multidimensional NMR spectroscopy (e.g. COSY, RELAY, TOCSY, NOESY, ROESY) is introduced. Isotope labelling and spectral editing strategies (e.g. HSQC, HMBC, TOCSY-HSQC, HNCO, HNCA) are outlined. Basics of NMR structure calculation will be described, including data collection, resonance assignment, collection of structural restraints, target function minimization and quality assessment. Different approaches related to structure determination of proteins (smaller than 15 kDa, 15-30 KDa and larger than 30 kDa) are described.
Comparison of NMR- and X-ray determined 3D-structures are discussed. Protein folding is introduced viewed by NMR spectroscopy: namely thermal unfolding, hydrogen-deuterium exchange, intrinsically unstructured proteins etc.. Examples regarding enzymes and their function, seen by NMR spectroscopy will be provided. NMR spectroscopy of nucleic acids and carbohydrates will also be introduced briefly.
Do not hesitate to come: this will be a lifetime experience, you will like it and understand NMR! (Language is optional, English or Hungarian upon request.)