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
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/572465
- author
- 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.
- organization
- publishing date
- 2007
- 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}}, }