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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
University of Toronto, ON, Canada.
Lund University, Sweden .
Lund University, Sweden .
University of Toronto, ON, Canada.
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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
Abstract [en]

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.
Keyword [en]
Selective 13C labeling, Protein expression, 1-13C]-glucose, [2-13C]-glucose, 13C relaxation measurements, CPMG relaxation dispersion, T1ρ
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:liu:diva-85031DOI: 10.1007/s10858-007-9158-6OAI: oai:DiVA.org:liu-85031DiVA: diva2:563719
Available from: 2012-10-31 Created: 2012-10-31 Last updated: 2017-12-07

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Lundström, Patrik

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CiteExportLink to record
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  • apa
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  • Other style
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  • de-DE
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  • sv-SE
  • Other locale
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