title: A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins.
abstract: A new method for stereospecific assignment of prochiral methyl groups in proteins is presented in which protein samples are produced using U-[(13)C]glucose and subsaturating amounts of 2-[(13)C]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 (1)H-(13)C 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.
authors.affiliation: CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France. michael.plevin@ibs.fr
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Plevin
authors.forename: Olivier
authors.initials: O
authors.lastname: Hamelin
authors.forename: Jérôme
authors.initials: J
authors.lastname: Boisbouvier
authors.forename: Pierre
authors.initials: P
authors.lastname: Gans
mesh_terms: D007553:Isotope Labeling; D007773:Lactates / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
chemical_list: D007773:Lactates; D011506:Proteins; C006359:alpha-acetolactate