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NESG Wiki: NMR Data Acquisition

NESG Wiki: NMR Data Acquisition. Alexander Eletsky 01/10/2010. Introduction. Documenting NESG protocols: reference and educational information Specific details for Varian and Bruker instruments Some expertise in NMR expected from users. Spectromer Operation.

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NESG Wiki: NMR Data Acquisition

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  1. NESG Wiki: NMR Data Acquisition Alexander Eletsky01/10/2010

  2. Introduction • Documenting NESG protocols: reference and educational information • Specific details for Varian and Bruker instruments • Some expertise in NMR expected from users HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  3. Spectromer Operation HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  4. NMR Experiments for Structure Determination • Standard experiment sets for high-throughput NMR • Additional experiments and novel methods HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  5. “Standard” NMR Experiments HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  6. Stereospecific Assignment of Leu/Val Isopropyl Groups • Optimized CT [13C,1H]-HSQC (28ms, 42ms and 56ms) for fractionally 13C-labeled samples D. Neri, T. Szyperski, G. Otting, H. Senn, K. Wuethrich, Biochemistry1989; 28(19):7510-7516 HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  7. HistidineProtonation State • Long-range [15N,1H]-HSQC or HMQC • Also yields He1/Ne1 spin assignments J. G. Pelton, D. A. Torchia, N. D. Meadow, and S. Roseman, Protein Sci. 1993; 2(4): 543–558. HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  8. 3D 13C/15N-resolved NOESY • Optimized for simultaneous 13C(aliphatic), 13C(aromatic) and 15N frequency labeling • Uses adiabatic 13C decoupling sideband suppression (SEAD – E. Kupce) • Spectra acquired at 900 MHz • Three-fold time saving compared to separate experiments S. M. Pascal, D. R. Muhandiram, T. Yamazaki, J. D. Forman-Kay, and L. E. Kay, JMR Ser. B. 1994; 103:197–201.Y. Shen, H. S. Atreya, G. Liu, and T. Szyperski, JACS 2005; 127:9085-9099. HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

  9. Acknowledgements • James M. Aramini • AleksandrasGutmanas • Gaohua Liu • Jeffrey L. Mills • Theresa A. Ramelot • Paolo Rossi • Kiran K. Singarapu • Yibing Wu • DuanxiangXu • Qi Zhang HTP Methods for Protein NMR Structure Analysis Used by the NIGMS Protein Structure initiative, January 10, 2010

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