Biomedical Engineering Reference
In-Depth Information
Figure 2.10
Nitrogen-detected CAN and CON experiments recorded with 52 kDa
GST. (A and B) Nitrogen-detected (A) CAN and (B) CON spectra of a
uniformly
2
H
15
N
13
C-labelled GST dimer recorded at 298 K in buffered
D
2
O supplemented with 4 mM Gd (DTPA-BMA). The dashed rectangle
in (A) contains the proline C
d
-N correlations (top left). The proline C
d
-
N correlations are connected to C
a
-N correlations as well as C9-N
correlations in (B) by cyan lines. (C) Proton-detected
15
N-
1
H TROSY
spectrum recorded on the same sample without Gd (DTPA-BMA).
Adapted from ref 41.
in NCO-IPAP, which use the 'out-and-back' coherence pathway. Thus, the
CON experiment may be less susceptible to signal loss due to CSA at high
molecular weights and in high magnetic fields. Figure 2.10(B) shows the CON
spectrum for the 52 kDa GST protein dimer. The spectrum was recorded over
1.5 days, and we observed y160 resolved signals, which correspond to y71%
of the expected resonances.
2.6 Low-c Detection in Interaction Studies and
Structure Determination
Obtaining structure information from low-c nuclei detection might have some
limitations due to the sparse spin-spin interactions. It has been shown that the
