Fractional C-13 enrichment of isolated carbons using [1-C-13]- or [2-C-13]-glucose facilitates the accurate measurement of dynamics at backbone C-alpha and side-chain methyl positions in proteins
2007 (English)In: Journal of Biomolecular NMR, ISSN 0925-2738, E-ISSN 1573-5001, Vol. 38, no 3, 199-212 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
Springer, 2007. Vol. 38, no 3, 199-212 p.
Selective 13C labeling, Protein expression, 1-13C]-glucose, [2-13C]-glucose, 13C relaxation measurements, CPMG relaxation dispersion, T1ρ
IdentifiersURN: urn:nbn:se:liu:diva-85031DOI: 10.1007/s10858-007-9158-6OAI: oai:DiVA.org:liu-85031DiVA: diva2:563719