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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 orcid 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
; ; ; ; ; ; ; and
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
2016-04-01 12:27:05
date last changed
2024-06-11 11:10:14
@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}},
  keywords     = {{[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}},
  doi          = {{10.1007/s10858-007-9158-6}},
  volume       = {{38}},
  year         = {{2007}},
}