Determination of the three-dimensional solution structure of the histidine-containing phosphocarrier protein HPr from Escherichia coli using multidimensional NMR spectroscopy.

TitleDetermination of the three-dimensional solution structure of the histidine-containing phosphocarrier protein HPr from Escherichia coli using multidimensional NMR spectroscopy.
Publication TypeJournal Article
Year of Publication1992
Authorsvan Nuland, N. A. J., J. Grötzinger, K. Dijkstra, R. M. Scheek, and G. T. Robillard
JournalEur J Biochem
Volume210
Issue3
Pagination881-91
Date Published1992 Dec 15
ISSN0014-2956
KeywordsAmino 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 JournalEur. J. Biochem.
PubMed ID1483471