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Fractional C-13 enr...
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
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- Lundström, Patrik, 1971- (författare)
- University of Toronto, ON, Canada
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- Teilum, Kaare (författare)
- Lund University, Sweden
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- Carstensen, Tommy (författare)
- Lund University, Sweden
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- Bezsonova, Irina (författare)
- University of Toronto, ON, Canada
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- Wiesner, Silke (författare)
- University of Toronto, ON, Canada
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- Hansen, D. Flemming (författare)
- University of Toronto, ON, Canada
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- Religa, Tomasz L. (författare)
- MRC, University of Cambridge, UK
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- Akke, Mikael (författare)
- Lund University, Sweden
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- Kay, Lewis E. (författare)
- University of Toronto, ON, Canada
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(creator_code:org_t)
- 2007-06-07
- 2007
- Engelska.
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Ingår i: Journal of Biomolecular NMR. - : Springer. - 0925-2738 .- 1573-5001. ; 38:3, s. 199-212
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- 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.
Nyckelord
- Selective 13C labeling
- Protein expression
- 1-13C]-glucose
- [2-13C]-glucose
- 13C relaxation measurements
- CPMG relaxation dispersion
- T1ρ
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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