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Specific (CD2CD2SCHD2)-C-12-D-beta-C-12-D-gamma-C-13-H-epsilon Isotopomer Labeling of Methionine To Characterize Protein Dynamics by H-1 and C-13 NMR Relaxation Dispersion

Weininger, Ulrich LU ; Liu, Zhihong; McIntyre, Deane D.; Vogel, Hans J. and Akke, Mikael LU (2012) In Journal of the American Chemical Society 134(45). p.18562-18565
Abstract
Protein dynamics on the micro- to millisecond time scale is increasingly found to be critical for biological function, as demonstrated by numerous NMR relaxation dispersion studies. Methyl groups are excellent probes of protein interactions and dynamics because of their favorable NMR relaxation properties, which lead to sharp signals in the H-1 and C-13 NMR spectra. Out of the six different methyl-bearing amino acid residue types in proteins; methionine plays a special role because of its extensive side chain flexibility and the high polarizability of the sulfur atom. Methionine is over-represented:in many protein-protein recognition sites making the methyl group of this, residue type an important probe of the relationships among dynamics,... (More)
Protein dynamics on the micro- to millisecond time scale is increasingly found to be critical for biological function, as demonstrated by numerous NMR relaxation dispersion studies. Methyl groups are excellent probes of protein interactions and dynamics because of their favorable NMR relaxation properties, which lead to sharp signals in the H-1 and C-13 NMR spectra. Out of the six different methyl-bearing amino acid residue types in proteins; methionine plays a special role because of its extensive side chain flexibility and the high polarizability of the sulfur atom. Methionine is over-represented:in many protein-protein recognition sites making the methyl group of this, residue type an important probe of the relationships among dynamics, Interactions, and biological function. Here we present a straightforward method to label methionine residues with specific (CHD2)-C-13 methyl isotopomers against a deuterated background. The resulting protein samples yield NMR spectra with improved sensitivity due to the essentially 100% population of the desired (CHD2)-C-13 methyl isotopomer, which is ideal for H-1 and C-13 spin relaxation experiments to investigate protein dynamics in general and conformational exchange in particular. We demonstrate the approach by measuring H-1 and C-13 CPMG relaxation dispersion for the nine methionines in calcium free calmodulin (apo-CaM). The results. show that the C- terminal domain, but not the N-terminal domain, of apo-CaM undergoes fast exchange between the ground state and a high-energy state. Since target proteins are known to bind specifically to the C-terminal domain of apo-CaM, we speculate that the high energy state might be involved in target binding through conformational selection. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of the American Chemical Society
volume
134
issue
45
pages
18562 - 18565
publisher
The American Chemical Society
external identifiers
  • wos:000311192100022
  • scopus:84869381828
ISSN
1520-5126
DOI
10.1021/ja309294u
language
English
LU publication?
yes
id
726e6466-77ff-4b25-8cc1-529d257f79bf (old id 3388242)
date added to LUP
2013-01-30 14:17:20
date last changed
2017-10-22 04:07:22
@article{726e6466-77ff-4b25-8cc1-529d257f79bf,
  abstract     = {Protein dynamics on the micro- to millisecond time scale is increasingly found to be critical for biological function, as demonstrated by numerous NMR relaxation dispersion studies. Methyl groups are excellent probes of protein interactions and dynamics because of their favorable NMR relaxation properties, which lead to sharp signals in the H-1 and C-13 NMR spectra. Out of the six different methyl-bearing amino acid residue types in proteins; methionine plays a special role because of its extensive side chain flexibility and the high polarizability of the sulfur atom. Methionine is over-represented:in many protein-protein recognition sites making the methyl group of this, residue type an important probe of the relationships among dynamics, Interactions, and biological function. Here we present a straightforward method to label methionine residues with specific (CHD2)-C-13 methyl isotopomers against a deuterated background. The resulting protein samples yield NMR spectra with improved sensitivity due to the essentially 100% population of the desired (CHD2)-C-13 methyl isotopomer, which is ideal for H-1 and C-13 spin relaxation experiments to investigate protein dynamics in general and conformational exchange in particular. We demonstrate the approach by measuring H-1 and C-13 CPMG relaxation dispersion for the nine methionines in calcium free calmodulin (apo-CaM). The results. show that the C- terminal domain, but not the N-terminal domain, of apo-CaM undergoes fast exchange between the ground state and a high-energy state. Since target proteins are known to bind specifically to the C-terminal domain of apo-CaM, we speculate that the high energy state might be involved in target binding through conformational selection.},
  author       = {Weininger, Ulrich and Liu, Zhihong and McIntyre, Deane D. and Vogel, Hans J. and Akke, Mikael},
  issn         = {1520-5126},
  language     = {eng},
  number       = {45},
  pages        = {18562--18565},
  publisher    = {The American Chemical Society},
  series       = {Journal of the American Chemical Society},
  title        = {Specific (CD2CD2SCHD2)-C-12-D-beta-C-12-D-gamma-C-13-H-epsilon Isotopomer Labeling of Methionine To Characterize Protein Dynamics by H-1 and C-13 NMR Relaxation Dispersion},
  url          = {http://dx.doi.org/10.1021/ja309294u},
  volume       = {134},
  year         = {2012},
}