Comparisons of NMR Spectral Quality and Success in Crystallization Demonstrate that NMR and X-ray Crystallography Are Complementary Methods for Small Protein Structure Determination

J. Am. Chem. Soc., 127 (47), 16505 -16511, 2005. 10.1021/ja053564h S0002-7863(05)03564-X
Web Release Date: November 8, 2005

Comparisons of NMR Spectral Quality and Success in Crystallization Demonstrate that NMR and X-ray Crystallography Are Complementary Methods for Small Protein Structure Determination

David A. Snyder, Yang Chen, Natalia G. Denissova, Thomas Acton, James M. Aramini, Melissa Ciano, Richard Karlin, Jinfeng Liu, Philip Manor, P. A. Rajan, Paolo Rossi, G. V. T. Swapna, Rong Xiao, Burkhard Rost, John Hunt,* and Gaetano T. Montelione*

Contribution from the Center for Advanced Biotechnology and Medicine, Department of Molecular Biology and Biochemistry, and Northeast Structural Genomics Consortium, Rutgers University, Piscataway, New Jersey 08854, Department of Biological Sciences, and Northeast Structural Genomics Consortium, Columbia University, New York, New York 10027, Department of Biochemistry and Molecular Biophysics, and Northeast Structural Genomics Consortium, Columbia University, 650 West 168th Street BB217, New York, New York 10032, and Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854

guy@cabm.rutgers.edu; jfhunt@biology.columbia.edu

Received May 31, 2005

Abstract:

X-ray crystallography and NMR spectroscopy provide the only sources of experimental data from which protein structures can be analyzed at high or even atomic resolution. The degree to which these methods complement each other as sources of structural knowledge is a matter of debate; it is often proposed that small proteins yielding high quality, readily analyzed NMR spectra are a subset of those that readily yield strongly diffracting crystals. We have examined the correlation between NMR spectral quality and success in structure determination by X-ray crystallography for 159 prokaryotic and eukaryotic proteins, prescreened to avoid proteins providing polydisperse and/or aggregated samples. This study demonstrates that, across this protein sample set, the quality of a protein's [¹⁵N-¹H]-heteronuclear correlation (HSQC) spectrum recorded under conditions generally suitable for 3D structure determination by NMR, a key predictor of the ability to determine a structure by NMR, is not correlated with successful crystallization and structure determination by X-ray crystallography. These results, together with similar results of an independent study presented in the accompanying paper (Yee, et al., J. Am. Chem. Soc., accompanying paper), demonstrate that X-ray crystallography and NMR often provide complementary sources of structural data and that both methods are required in order to optimize success for as many targets as possible in large-scale structural proteomics efforts.

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