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Fractional 13C enrichment of isolated carbons using [1-13C]- or [2-13C]-glucose facilitates the accurate measurement of dynamics at backbone Ca and side-chain methyl positions in protein

Lundström, Patrik LU ; Teilum, Kaare LU ; Carstensen, Tommy; Bezsonova, Irina; Wiesner, Silke; Hansen, D. Flemming; Religa, Tomasz L.; Akke, Mikael LU and Kay, Lewis E. (2007) In Journal of Biomolecular NMR 38(3). p.199-212
Abstract
A simple labeling approach is presented based on protein expression in [1-C-13]- or [2-C-13]-glucose containing media that produces molecules enriched at methyl carbon positions or backbone C-alpha sites, respectively. All of the methyl groups, with the exception of Thr and Ile(delta 1) are produced with isolated C-13 spins (i.e., no C-13-C-13 one bond couplings), facilitating studies of dynamics through the use of spin-spin relaxation experiments without artifacts introduced by evolution due to large homonuclear scalar couplings. Carbon-alpha sites are labeled without concomitant labeling at C-beta positions for 17 of the common 20 amino acids and there are no cases for which C-13(alpha)-(CO)-C-13 spin pairs are observed. A large number... (More)
A simple labeling approach is presented based on protein expression in [1-C-13]- or [2-C-13]-glucose containing media that produces molecules enriched at methyl carbon positions or backbone C-alpha sites, respectively. All of the methyl groups, with the exception of Thr and Ile(delta 1) are produced with isolated C-13 spins (i.e., no C-13-C-13 one bond couplings), facilitating studies of dynamics through the use of spin-spin relaxation experiments without artifacts introduced by evolution due to large homonuclear scalar couplings. Carbon-alpha sites are labeled without concomitant labeling at C-beta positions for 17 of the common 20 amino acids and there are no cases for which C-13(alpha)-(CO)-C-13 spin pairs are observed. A large number of probes are thus available for the study of protein dynamics with the results obtained complimenting those from more traditional backbone N-15 studies. The utility of the labeling is established by recording C-13 R-1 rho and CPMG-based experiments on a number of different protein systems. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
[1-C-13]-glucose, protein expression, selective C-13 labeling, dispersion, CPMG relaxation, T1 rho, C-13 relaxation measurements, [2-C-13]-glucose
in
Journal of Biomolecular NMR
volume
38
issue
3
pages
199 - 212
publisher
Springer
external identifiers
  • wos:000247307400001
  • scopus:34250903564
ISSN
1573-5001
DOI
10.1007/s10858-007-9158-6
language
English
LU publication?
yes
id
ae5029b5-8db7-4b55-abe3-5443451c69ca (old id 572465)
date added to LUP
2007-11-05 08:56:12
date last changed
2017-10-22 03:53:37
@article{ae5029b5-8db7-4b55-abe3-5443451c69ca,
  abstract     = {A simple labeling approach is presented based on protein expression in [1-C-13]- or [2-C-13]-glucose containing media that produces molecules enriched at methyl carbon positions or backbone C-alpha sites, respectively. All of the methyl groups, with the exception of Thr and Ile(delta 1) are produced with isolated C-13 spins (i.e., no C-13-C-13 one bond couplings), facilitating studies of dynamics through the use of spin-spin relaxation experiments without artifacts introduced by evolution due to large homonuclear scalar couplings. Carbon-alpha sites are labeled without concomitant labeling at C-beta positions for 17 of the common 20 amino acids and there are no cases for which C-13(alpha)-(CO)-C-13 spin pairs are observed. A large number of probes are thus available for the study of protein dynamics with the results obtained complimenting those from more traditional backbone N-15 studies. The utility of the labeling is established by recording C-13 R-1 rho and CPMG-based experiments on a number of different protein systems.},
  author       = {Lundström, Patrik and Teilum, Kaare and Carstensen, Tommy and Bezsonova, Irina and Wiesner, Silke and Hansen, D. Flemming and Religa, Tomasz L. and Akke, Mikael and Kay, Lewis E.},
  issn         = {1573-5001},
  keyword      = {[1-C-13]-glucose,protein expression,selective C-13 labeling,dispersion,CPMG relaxation,T1 rho,C-13 relaxation measurements,[2-C-13]-glucose},
  language     = {eng},
  number       = {3},
  pages        = {199--212},
  publisher    = {Springer},
  series       = {Journal of Biomolecular NMR},
  title        = {Fractional 13C enrichment of isolated carbons using [1-13C]- or [2-13C]-glucose facilitates the accurate measurement of dynamics at backbone Ca and side-chain methyl positions in protein},
  url          = {http://dx.doi.org/10.1007/s10858-007-9158-6},
  volume       = {38},
  year         = {2007},
}