solution protein nmr - rieko ishima1h-15n nmr correlation spectra of hiv-1 protease once signals are...
TRANSCRIPT
Solution Protein NMR
Rieko Ishima Dept. of Structural Biology
School of Medicine
Nuclear Magnetic Resonance (NMR)
Nuclear Magnetic Resonance (NMR) spectroscopy
NMR spectroscopy - Resonance
B0
N
S
1H nuclear spin
Hydrogen atom
α
β
NMR spectroscopy - Resonance
α
β
Chemical shift (ppm) ηω
Nuclear spin
B0: Magnetic field strength
α
β hωΗ
(ppm)
At 14 Tesla Magnet Proton resonance frequency (ωH) = 600 MHz Carbon resonance frequency (ωC) = 150 MHz Nitrogen resonance frequency (ωH) = 61 MHz
NMR Spectroscopy - Resonance
Correlation
H H-N
N
ppm H
Observation of N-H chemical shifts
H C
CH H3C CH3
C O N H
C H
CH3 O C N
H
H C
CH NH2
C O
OC
H N
N - H
1H (ppm)
15N
(ppm
)
An N-H correlation spectrum
1H (ppm)
15N
(ppm
)
1H
15N
ωN
ωH
Three-dimensional NMR spectra
1H
13C
1H
15N
13C
Three-dimensional NMR spectra
1H
13C
1H
15N
13C
Backbone signal assignments CBCACONH
HNCA
Cαi+1
1H (ppm)
15N (ppm
)
T96
1H-15N NMR correlation spectra of HIV-1 protease Once signals are assigned,
the information is used to identify the sides that are affected by a mutation.
1H (ppm)
Dimer (PR) Monomer (PRR87K)
Ishima et al (2001) J Biol Chem. 276, 49110-6.