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1D NMR Spectrum of a Protein. AV-COSY. 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. 2D NOESY Spectrum of a Protein. Some Useful 2D NMR Experiments.
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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
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
IR24 FGF-2 JR19 OP3 LC8 WR33 Z-domain WR41 MMP-1 ZR18 ER115 N-TmZip WR64 ZR31 C-TmZip WR90 ER14 Protein NMR Structures IL13 ER75
C C N N ER75: Ecol YgdK (nmr)ER30: Ecol SufE (xray)IR24: Hinf IscU (nmr)SR17 Bsub IscU (nmr) YgdK cf SufE A. Kuzin et al. (Hunt) YgdK G. Liu et al. (Szyperski) IscU, SufE,YgdK Domain Families are Structurally Similar Gram pos IscU G. Kornhaber et al. (Montelione) Ramelot et al, J. Mol. Biol. 2004 Goldsmith et al., J. Mol. Biol. 2004 Liu et al. submitted Gram neg IscU T. Ramelot et al. (Kennedy)