Title | Determination of the three-dimensional solution structure of the histidine-containing phosphocarrier protein HPr from Escherichia coli using multidimensional NMR spectroscopy. |
Publication Type | Journal Article |
Year of Publication | 1992 |
Authors | van Nuland, N. A. J., J. Grötzinger, K. Dijkstra, R. M. Scheek, and G. T. Robillard |
Journal | Eur J Biochem |
Volume | 210 |
Issue | 3 |
Pagination | 881-91 |
Date Published | 1992 Dec 15 |
ISSN | 0014-2956 |
Keywords | Amino Acid Sequence, Bacterial Proteins, Carbon Isotopes, Escherichia coli, Hydrogen, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Sequence Data, Nitrogen Isotopes, Phosphoenolpyruvate Sugar Phosphotransferase System, Protein Conformation, Protein Folding |
Abstract | We recorded several types of heteronuclear three-dimensional (3D) NMR spectra on 15N-enriched and 13C/15N-enriched histidine-containing phosphocarrier protein, HPr, to extend the backbone assignments [van Nuland, N. A. J., van Dijk, A. A., Dijkstra, K., van Hoesel, F. H. J., Scheek, R. M. & Robillard, G. T. (1992) Eur. J. Biochem, 203, 483-491] to the side-chain 1H,15N and 13C resonances. From both 3D heteronuclear 1H-NOE 1H-13C and 1H-NOE 1H-15N multiple-quantum coherence (3D-NOESY-HMQC) and two-dimensional (2D) homonuclear NOE spectra, more than 1200 NOE were identified and used in a step-wise structure refinement process using distance geometry and restrained molecular dynamics involving a number of new features. A cluster of nine structures, each satisfying the set of NOE restraints, resulted from this procedure. The average root-mean-square positional difference for the C alpha atoms is less than 0.12 nm. The secondary structure topology of the molecule is that of an open-face beta sandwich formed by four antiparallel beta strands packed against three alpha helices, resembling the recently published structure of Bacillus subtilis HPr, determined by X-ray crystallography [Herzberg, O., Reddy, P., Sutrina, S., Saier, M. H., Reizer, J. & Kapafia, G. (1992) Proc. Natl, Acad. Sci. USA 89, 2499-2503). |
Alternate Journal | Eur. J. Biochem. |
PubMed ID | 1483471 |
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