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  A straightforward method for stereospecific assignment of val and leu prochiral methyl groups by solid-state NMR: Scrambling in the [2-13C]Glucose labeling scheme.

Lv, G., Faßhuber, H. K., Loquet, A., Demers, J. P., Vijayan, V., Giller, K., et al. (2013). A straightforward method for stereospecific assignment of val and leu prochiral methyl groups by solid-state NMR: Scrambling in the [2-13C]Glucose labeling scheme. Journal of Magnetic Resonance, 228, 45-49. doi:10.1016/j.jmr.2012.12.017.

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 Creators:
Lv, G.1, Author           
Faßhuber, H. K.2, Author           
Loquet, A.2, Author           
Demers, J. P.2, Author           
Vijayan, V.2, Author           
Giller, K.1, Author           
Becker, S.1, Author           
Lange, A.2, Author           
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1Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              
2Research Group of Solid-State NMR, MPI for Biophysical Chemistry, Max Planck Society, ou_persistent35              

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 Abstract: The unambiguous stereospecific assignment of the prochiral methyl groups in Val and Leu plays an important role in the structural investigation of proteins by NMR. Here, we present a straightforward method for their stereospecific solid-state NMR assignment based on [2-13C]Glucose ([2-13C]Glc) as the sole carbon source during protein expression. The approach is fundamentally based on the stereo-selective biosynthetic pathway of Val and Leu, and the co-presence of [2-13C]pyruvate produced mainly by glycolysis and [3-13C]/[1,3-13C]pyruvate most probably formed through scrambling in the pentose phosphate pathway. As a consequence, the isotope spin pairs 13Cβ-13Cγ2 and 13Cα-13Cγ1 in Val, and 13Cγ-13Cδ2 and 13Cβ-13Cδ1 in Leu are obtained. The approach is successfully demonstrated with the stereospecific assignment of the methyl groups of Val and Leu of type 3 secretion system PrgI needles and microcrystalline ubiquitin.

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Language(s): eng - English
 Dates: 2013-01-042013-03
 Publication Status: Issued
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 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.jmr.2012.12.017
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Title: Journal of Magnetic Resonance
Source Genre: Journal
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Pages: - Volume / Issue: 228 Sequence Number: - Start / End Page: 45 - 49 Identifier: ISSN: 0022-2364