[nmrlearner]

Speeding up direct 15N detection: hCaN 2D NMR experiment


Abstract Experiments detecting low gyromagnetic nuclei have recently been proposed to utilize the relatively slow relaxation properties of these nuclei in comparison to 1H. Here we present a new type of 15N direct-detection experiment. Like the previously proposed CaN experiment (Takeuchi et al. in J Biomol NMR 47:271â??282, 2010), the hCaN experiment described here sequentially connects amide 15N resonances, but utilizes the initial high polarization and the faster recovery of the 1H nucleus to shorten the recycling delay. This allows recording 2D 15N-detected NMR experiments on proteins within a few hours, while still obtaining superior resolution for 13C and 15N, establishing sequential assignments through prolines, and at conditions where amide protons exchange rapidly. The experiments are demonstrated on various biomolecules, including the small globular protein GB1, the 22 kDa HEAT2 domain of eIF4G, and an unstructured polypeptide fragment of NFAT1, which contains many SerPro sequence repeats.

• Content Type Journal Article
• Category Article
• Pages 1-8
• DOI 10.1007/s10858-011-9580-7
• Authors
  • Maayan Gal, Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA
  • Katherine A. Edmonds, Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA
  • Alexander G. Milbradt, Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA
  • Koh Takeuchi, Biomedicinal Information Research Center, National Institute of Advanced Industrial Science and Technology, Tokyo, 135-0064 Japan
  • Gerhard Wagner, Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA

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

Source: Journal of Biomolecular NMR