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

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-[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.

• Content Type Journal Article
• Pages 1-7
• DOI 10.1007/s10858-010-9463-3
• Authors
  • Michael J. Plevin, CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
  • Olivier Hamelin, CNRS, Laboratoire de Chimie et Biologie des Métaux, 17 avenue des martyrs, Grenoble, France
  • Jérôme Boisbouvier, CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
  • Pierre Gans, CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR