A spectral correlation function for efficient sequential NMR assignments of uniformly 15N-labeled proteins |
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Authors: | Christian Bartels Kurt Wüthrich |
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Institution: | (1) Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule-Hönggerberg, CH-8093 Zürich, Switzerland |
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Abstract: | Summary A new computer-based approach is described for efficient sequence-specific assignment of uniformly 15N-labeled proteins. For this purpose three-dimensional 15N-correlated 1H, 1H]-NOESY spectra are divided up into two-dimensional 1H-1H strips which extend over the entire spectral width along one dimension and have a width of ca. 100 Hz, centered about the amide proton chemical shifts along the other dimension. A spectral correlation function enables sorting of these strips according to proximity of the corresponding residues in the amino acid sequence. Thereby, starting from a given strip in the spectrum, the probability of its corresponding to the C-terminal neighboring residue is calculated for all other strips from the similarity of their peak patterns with a pattern predicted for the sequentially adjoining residue, as manifested in the scalar product of the vectors representing the predicted and measured peak patterns. Tests with five different proteins containing both -helices and -sheets, and ranging in size from 58 to 165 amino acid residues show that the discrimination achieved between the sequentially neighboring residue and all other residues compares well with that obtained with an unguided interactive search of pairs of sequentially neighboring strips, with important savings in the time needed for complete analysis of 3D 15N-correlated 1H, 1H]-NOESY spectra. The integration of this routine into the program package XEASY ensures that remaining ambiguities can be resolved by visual inspection of the strips, combined with reference to the amino acid sequence and information on spin-system types obtained from additional NMR spectra.Abbreviations 1D, 2D, 3D, 4D
one-, two-, three-, four-dimensional
- NOE
nuclear Overhauser enhancement
- NOESY
nuclear Overhauser enhancement spectroscopy
- COSY
correlation spectroscopy
- TOCSY
total correlation spectroscopy |
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Keywords: | Proteins Sequential resonance assignment Interactive software XEASY |
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