1. Solution Structure Determination by NMR and Distance Geometry Calculations 1. Pure Protein (0.3 mL, 0.5 - 1 mM; ~ 10 mg) 2. Sequence-specific resonance assignments 3. Sequence-specifice distance constraints disulfide bonds spin-spin coupling constants internuclear distance constraints from NOEs residual dipolar couplings hydrogen bonds standard bond lengths; bond angles 4. Distance Geometry Calculations 5. Constrained Energy Minimization; Refinement

2. Basic Principles Nuclear Spin and Magnetization

4. Magnetization and Coherence

6. Fourier Transform NMR Ernst & Anderson Rev. Scientific Instrumentation 37: 93 (1966)

8. Nuclear Relaxation

10. Spin-Spin Coupling

11. Chemical Shift with Spin-Spin Coupling

12. 1D NMR Spectrum of a Protein

13. 2D Correlated SpectroscopY

14. AV-COSY

15. Nuclear Overhauser Effect (NOE) Def: The change in the intensity of resonance i upon irradiation of resonance j due to “cross relaxation” between spins i and j through space

17. 2D NOESY Spectrum of a Protein

18. Some Useful 2D NMR Experiments

19. 1 H- 13 C Heteronuclear Correlation Spectrum

21. Solution Structure Determination by NMR and Distance Geometry Calculations 1. Pure Protein (0.3 mL, 0.5 - 1 mM; ~ 10 mg) 2. Sequence-specific resonance assignments 3. Sequence-specifice distance constraints disulfide bonds spin-spin coupling constants internuclear distance constraints from NOEs residual dipolar couplings hydrogen bonds standard bond lengths; bond angles 4. Distance Geometry Calculations 5. Constrained Energy Minimization; Refinement

25. WR41 Protein NMR Structures C-TmZip ER14 MMP-1 IL13 FGF-2 WR90 WR64 LC8 ER115 N-TmZip JR19 ZR18 OP3 WR33 ZR31 ER75 Z-domain IR24

26. ER75: Ecol YgdK (nmr) ER30: Ecol SufE (xray) IR24: Hinf IscU (nmr) SR17 Bsub IscU (nmr) N N CC YgdK G. Liu et al. (Szyperski) Gram neg IscU IscU, SufE,YgdK Domain Families are Structurally Similar YgdK cf SufE A. Kuzin et al. (Hunt) Ramelot et al, J. Mol. Biol. 2004 Goldsmith et al., J. Mol. Biol. 2004 Liu et al. submitted T. Ramelot et al. (Kennedy) Gram pos IscU G. Kornhaber et al. (Montelione)