A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins

Michael J Plevin (1) , Olivier Hamelin (2) , Jérôme Boisbouvier (1) , Pierre Gans (1)

1 IBS - UMR 5075 - Institut de biologie structurale
2 LCBM - UMR 5249 - Laboratoire de Chimie et Biologie des Métaux

A new method for stereospecific assignment of prochiral methyl groups in proteins is presented in which protein samples are produced using U-[13C]glucose and subsaturating amounts of 2-[13C]methyl-acetolactate. The resulting non-uniform labeling pattern allows proR and proS methyl groups to be easily distinguished by their different phases in a constant-time two-dimensional 1H-13C correlation spectra. Protein samples are conveniently prepared using the same media composition as the main uniformly-labeled sample and contain higher levels of isotope-enrichment than fractional labeling approaches. This new strategy thus represents an economically-attractive, robust alternative for obtaining isotopically-encoded stereospecific NMR assignments of prochiral methyl groups.

Mots clés: MALATE-SYNTHASE-G; MOLECULAR-WEIGHT PROTEINS; NUCLEAR-MAGNETIC-RESONANCE; Prochiral methyl groups; Acetolactate; Stereospecific assignments; Isotope labeling; NMR ASSIGNMENTS; C-13; LEUCINE; VAL; LEU; RESIDUES; SPECTROSCOPY

Domaines: Biologie structurale [q-bio.BM]

Michael J Plevin, Olivier Hamelin, Jérôme Boisbouvier, Pierre Gans. A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins. Journal of Biomolecular NMR, 2011, 49 (2), pp.61-67. ⟨10.1007/s10858-010-9463-3⟩. ⟨hal-01138581⟩