1. -A cell is an organization of millions of molecules -Proper communication between these molecules is essential to the normal functioning of the cell -To understand communication between molecules *determine the arrangement of the atoms* *Atomic Resolution Structural Biology* Organ  Tissue  Cell  Molecule  Atoms Medicine and Biology at the Atomic Scale

2. High Resolution Structural Biology Using atomic structure to understand how biomolecules communicate

3. Anticancer Drug Duocarmycin SA Biological Specificity from Structure Shape Atomic interactions

4. Putting Structures in Context Molecule Structural Genomics Pathway Structural Proteomics Activity Systems Biology RPA NER BER RR

5. Strategy and Approaches NMR Spectroscopy X-ray Crystallography Computation It’s not about the structure or the technique, it’s about the Biology!!!!

6. Determination of 3D Structures X-rayNMR X-ray Diffraction Pattern RF Resonance H0H0  Direct detection of atom positions  Crystals  Indirect detection of H-H distances  In solution

7. Static Structure/Dynamic Biology Structures determined by NMR, computation, and X-ray crystallography are static snapshots of highly dynamic molecular systems Biological process (recognition, interaction, chemistry) require molecular motions (from femto-seconds to minutes) New methods are needed to comprehend and facilitate thinking about the dynamic structure of molecules: visualization

8. Challenges for Converting Structure to Function Neither crystal nor solution structures are properly represented by a single low energy conformer To be called a structure (not a model), there must be a complete representation of what parts are well known and what parts are not well known

9. Conformational Ensemble: Representing a Structure C N Uncertainty/Precision: RMSD of the ensemble

10. Unique Contributions X-ray crystallography- highest resolution structures; fast NMR- enables widely varying solution conditions; characterization of motions and dynamic, weakly interacting systems Computation- fundamental understanding of structure, dynamics and interactions; models without experiment

11. Center for Structural Biology Increase molecular thinking Foster multi-disciplinary research Focal point bridging medicine and biology to math, physics, chemistry >12 research groups (8 new!!) Promote collaborations involving structural biology researchPromote collaborations involving structural biology research

12. How Does NMR Work? HoHo parallel anti-parallel Alignment Energy  E = h  H o Efficiency factor- nucleus ConstantsStrength of magnet

13. Resonance: Perturb Equilibrium ap h  E p ap p equilibrium non-equilibrium If energy is input at exactly the right frequency ( ), it gets absorbed EE

14. Resonance: Read Out Signals +-+- H o = +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- +-+- 1. force non-alignment2. release p ap p p

15. NMR of Molecules CH 3 -CH 2 -OH Resonance assignment: which signal for which H atoms? OHCH 2 CH 3 10 0

16. Challenges of NMR on Proteins Proteins have hundreds/thousands of signals Resonance assignment is a huge challenge Need to use computer programs to convert from NMR data to structures

17. Uses of Solution NMR to Elucidate Protein Function Analytical and Physical Characterization –Is the protein folded? –Sequence, post-translational modifications –Important: measure binding constants Assess structural homology/effect of mutations Three-dimensional structure determination Unique: Measure flexibility/dynamicsUnique: Measure flexibility/dynamics *To address “static structure/dynamic biology”*