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1D NMR Spectrum of a Protein - PowerPoint PPT Presentation


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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
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

slide13

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

er75 ecol ygdk nmr er30 ecol sufe xray ir24 hinf iscu nmr sr17 bsub iscu nmr

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)