caa a22 4500 445325356 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s10858-011-9580-7 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9580-7 Speeding up direct 15N detection: hCaN 2D NMR experiment [Elektronische Daten] [Maayan Gal, Katherine Edmonds, Alexander Milbradt, Koh Takeuchi, Gerhard Wagner] 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 47:271-282, 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 22kDa HEAT2 domain of eIF4G, and an unstructured polypeptide fragment of NFAT1, which contains many SerPro sequence repeats. Springer Science+Business Media B.V., 2011 15N direct detection nationallicence CaN nationallicence hCaN nationallicence Nuclear magnetic resonance (NMR) nationallicence Gal Maayan Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut Edmonds Katherine Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut Milbradt Alexander Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut Takeuchi Koh Biomedicinal Information Research Center, National Institute of Advanced Industrial Science and Technology, 135-0064, Tokyo, Japan aut Wagner Gerhard Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut Journal of Biomolecular NMR Springer Netherlands 51/4(2011-12-01), 497-504 0925-2738 51:4<497 2011 51 10858 https://doi.org/10.1007/s10858-011-9580-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9580-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Gal Maayan Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut NATIONALLICENCE 700 1- Edmonds Katherine Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut NATIONALLICENCE 700 1- Milbradt Alexander Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut NATIONALLICENCE 700 1- Takeuchi Koh Biomedicinal Information Research Center, National Institute of Advanced Industrial Science and Technology, 135-0064, Tokyo, Japan aut NATIONALLICENCE 700 1- Wagner Gerhard Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, 02115, Boston, MA, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/4(2011-12-01), 497-504 0925-2738 51:4<497 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer