PRE-MEETING COURSE INTERMOLECULAR INTERACTIONS BY NMR MAY 01-02, 2009 - INSTITUTO MILITAR DE ENGENHARIA RIO DE JANEIRO, RJ, BRAZIL LECTURERS: EURICO J. S. CABRITA (PORTUGAL) FERNANDO LÓPEZ-ORTIZ (SPAIN) MARTA BRUIX (SPAIN) MIQUEL PONS (SPAIN)

MAY 1, 2009 - Methodology

MORNING

Fernando Ortiz and Eurico Cabrita - “INTRODUCTION. THEORY OF DIFFUSION AND NOE BASED EXPERIMENTS” - 4 HOURS

Topics Covered: Simple observations: effects of intermolecular interactions on chemical shift, heteronuclear coupling, relaxation, etc. Multinuclear Magnetic Resonance. (Fernando Ortiz, 1 hour); Measuring Diffusion by NMR: Concepts, pulse sequences and limitations (Eurico Cabrita, 1h 30 min); Concepts of NOE: Homonuclear NOE (Eurico Cabrita, 30 minutes); Heteronuclear NOE (Fernando Ortiz, 1h 00 min.).

AFTERNOON

Miquel Pons - “THEORY OF RELAXATION. STD AND PARAMAGNETIC PROBES” - 2 HOURS

Topics Covered: Principles of NMR Relaxation theory. Correlation time effects; Relaxation in chemically exchanging systems. Relaxation time scale; Detecting protein-protein interaction by changes in correlation times; Intermolecular Magnetization transfer; STD and WaterLOGSY; Spy molecules and protein-protein interactions; Epitope mapping; Kissing complexes: identification of binding interfaces; Paramagnetic probes and structural characterization of transient complexes.

Marta Bruix - “METHODOLOGY FOR THE 3D STRUCTURE DETERMINATION OF PROTEINS AND PROTEIN COMPLEXES” - 2 HOURS

Topics Covered: NMR parameters with structural information; Isotope labeling strategies, Isotope filtered and edited NMR methods for the study of biomolecular structure and interactions; Multidimensional heteronuclear assignment protocols, Extraction of structural restraints; Special NMR techniques for large molecular assemblies, deuteration, TROSY.


MAY 2, 2009 – APPLICATIONS

MORNING

Fernando Ortiz and Eurico Cabrita: “INTERMOLECULAR INTERACTIONS IN THE FIELD OF ORGANOMETALLIC CHEMISTRY AND SMALL MOLECULES” - 4 HOURS

Topics Covered: Diffusion NMR (Eurico Cabrita, 1h 30 m): Determination of aggregation and estimation of molecular weight; Binding studies and estimation of association constants; Diffusion studies using non-conventional nuclei. Applications in catalysis (effect of temperature, aggregation, ion-pair strength, etc). Applications of NOE (Fernando Ortiz, 1h 30 m): Homonuclear and heteronuclear NOE: applications (solute solvent interactions, ion-pairs, etc); Mechanistic studies and other Techniques (Fernando Ortiz and Eurico Cabrita, 1 hour); Para-hydrogen. Applications in coordination chemistry; Laser polarization. Applications in supramolecular Chemistry; HR-MAS. Applications in organic and organometallic chemistry (differences in mobility, surface interactions in solid phase supports, etc.); NMR in supercritical CO2 (high pressure NMR). Applications to the study of solute-solvent interactions.

AFTERNOON

Miquel Pons and Marta Bruix: “Structural studies of protein-ligand, protein-protein and protein-nucleic acid interactions by NMR” – 4 hours

Topics Covered: Miquel Pons - Weak complexes (2 hours): Case study 1- Oligomerization of low molecular weight phosphatases studied by 15N NMR relaxation, 129Xe NMR, Small Angle X-ray Scattering, and Docking; Case study 2- Examples of STD based characterization of small molecule protein complexes. Marta Bruix -Tight complexes (2 hours): Three dimensional structure determination, Methods based on NOE and RDC effects. Docking strategies; Structure of the ligand in the complex. Examples; Structure of the receptor, protein-protein and protein-nucleic acid complexes. Differences between the free and bound conformations. Examples.