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

作者：Plevin Michael J^*; Hamelin Olivier; Boi**ouvier Jerome; Gans Pierre

来源：Journal of Biomolecular NMR, 2011, 49(2): 61-67.

DOI：10.1007/s10858-010-9463-3

• Isotope labeling
• Stereospecific assignments
• Acetolactate
• Prochiral methyl groups

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.

• 出版日期2011-2
• 单位中国地震局