72-optimalizovany INADEQUATE
1
72-optimalizovaný INADEQUATE L
Korelace disperze chemických posunů L
2
Korelace disperze chemických posunů - disorder
Celulóza
(76.6)
C4 C3   i   C5 C6
iQ0.5|        (78.3i   (73.7) (64.71
C4 C5 C2
(88.2)        C3 1.74.5) (77.8)1 I
In more complex systems, correlations can be exploited to extract chains of correlated shifts. Each chain corresponds to the chemical shifts of an entire subunit with a given conformation.
Korelace disperze chemických posunů - deuterace
3
Korelace disperze chemických posunů - susceptibilita
Chemical Disorder (local effect) a change in the isotropic chemical shift
from one molecule to another Sj gj
e.g.. glasses, catalysts, polymers
Magnetic Susceptibility (bulk effect) a change in the Larmor frequency from one part of the sample to another
heterogeneous samples
How can the chemical shift differences
due to disorder be removed without removing the chemical shift differences that distinguish sites from each other?
--
Korelace disperze chemických posunů - refokusace
Zero-Quantum Spectroscopy
For susceptibility broadening or
broadening due to an inhomogeneous applied field, the shifts at one nucleus are perfectly correlated in a one-to-one fashion with the shifts at another nearby nucleus
Thus, while the individual chemical shifts change, the chemical shift differences remain the same
Hailetal. 1981:Zero-quantum NMR spectra are free of Bo broadening
Warren et ai. 1996: Intermolecular solvent-solute ZQ coherences yield high-resolution spectra in unstable very high magnetic fields
Terao etal. 1999: ZQ sideband spectra are free of susceptibility broadening in paramagnetic solids
4
Korelace disperze chemických posunů - refokusace
■
-►
Fli-ľ-sf-li-ľ-r-Mi-h-oi Cliŕnikŕil ::íiift ■■ |:-|:■ m
št* Fi"»i:|u*ni;y aril, units
To record this red spectrum we must refocus the chemical shift distribution due to disorder within a site, without refccusing the chemical shifts that distinguish sites from each other.
This cannot be done if the sites ere isolated from each other.
It can be achieved if
(ij the sites can be connected in a multi-dimensional spectrum
(ii) the chemical shifts between coupled pairs of spins are highly correlated.
Korelace disperze chemických posunů - refokusace
3ip_3ip Through-Bond Connectivities
-"'P bin:;ilř:-Qi.i:ir'itijn"i Frequency
Four in equivalent P-N-P pairs are revealed
1
The Reiocused INADEQUATE Experiment itai, JACS 121, 10987(1999)
5
Korelace disperze chemických posunů - refokusace
13C-13C korelace (PDSD) - dipolární interakce
Proton Driven Spin Diffusion
e
3D struktura prionových proteinů (2005) L
7
2D 13C-13C korelace - 1H-1H spinová difuse
H I t.. ■
t --	SPINAL	tmix	SPINAL	
	'>1		S4	
M4SCt M51 Ce
: v.. - 3 ms   :   i    i j
iBO  l*a  no  in)  mo   bo    so    «» 20 13C Chemical Shin (ppm)
5.D >.		á ■	
50 S46CU-E	«tu 1 T30,.9	Ir:	
	: j	/	IP                          > MICp.Ca j ^,S46Cp-Ca
70- M- T3GCP-G [ on T59.62CU-C	i		
isc Chemical Shift (ppm)
A. Böckmann, M. Juy, E. Bettler, L. Emsley, A. Galinier, F. Penin, A. Lesage .
Water-Protein Hydrogen Exchange in the Micro-Crystalline Protein Crh as Observed by Solid State NMR Spectroscopy, Journal of Biomolecular NMR , 32 195 (2005).
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2D 13C-15N dvojitá cross-polarizace L
[*" ' I      ti     I 1
Acquisition
t2
Avance 500 WB
4 mm sonda A 1H/15N, 2 ms kontakt A 15N/13C, 1 ms kontakt A variable amplitude
(50 % ramp) A 15N RF field: 64 kHz A 13C RF field: 64 kHz A 1H RF field: 89 kHz
- -240 -22o
200 150
HO
H:
l5N:
,JC:
- -300
-280
-260
- -200
100
50
ppm
Castellani, F., van Rossum, B.J., Diehl, A., Schubert, M., Rehbein, K., and Oschkinat, H. Structure of a protein determined by solid-state magic-angle-spinning NMR
Experimenty s dvojitou cross-polanzaci spectroscopy, Nature 420, 98-102 (2002).
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Tří-rezonanční experimenty 13C-15N korelace
Experimenty s dvojitou cross-polarizací Kostrukce sondy
Souhrn
Korelovaný disorder
L ň
13C-13C PDSD
Solid-state NMRK
■ and............. ■
-i-
Double-CP-13C-15N
3D Double CP-PDSD
■JtC
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