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Selective backbone labelling of ILV methyl labelled proteins
Ist Teil von
Journal of biomolecular NMR, 2009-04, Vol.43 (4), p.219-227
Ort / Verlag
Dordrecht: Dordrecht : Springer Netherlands
Erscheinungsjahr
2009
Link zum Volltext
Quelle
SpringerLink (Online service)
Beschreibungen/Notizen
Adding the ¹³C labelled 2-keto-isovalerate and 2-oxobutanoate precursors to a minimal medium composed of ¹²C labelled glucose instead of the commonly used (²D, ¹³C) glucose leads not only to the ¹³C labelling of (I, L, V) methyls but also to the selective ¹³C labelling of the backbone Cα and CO carbons of the Ile and Val residues. As a result, the backbone (¹H, ¹⁵N) correlations of the Ile and Val residues and their next neighbours in the (i + 1) position can be selectively identified in HN(CA) and HN(CO) planes. The availability of a selective HSQC spectrum corresponding to the sole amide resonances of the Ile and Val residues allows connecting them to their corresponding methyls by the intra-residue NOE effect, and should therefore be applicable to larger systems.