Open AccessDetermination of the three‐dimensional solution structure of the histidine‐containing phosphocarrier protein HPr from Escherichia coli using multidimensional NMR spectroscopy
Author(s) -
NULAND Nico A. J.,
GRÖTZINGER Joachim,
DIJKSTRA Klaas,
SCHEEK Ruud M.,
ROBILLARD George T.
Publication year - 1992
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1992.tb17492.x
Subject(s) - heteronuclear molecule , homonuclear molecule , antiparallel (mathematics) , chemistry , crystallography , two dimensional nuclear magnetic resonance spectroscopy , nuclear overhauser effect , stereochemistry , nuclear magnetic resonance spectroscopy , histidine , molecule , spectroscopy , physics , amino acid , biochemistry , organic chemistry , quantum mechanics , magnetic field
We recorded several types ofheteronuclear three‐dimensional (3D) NMR spectra on 15 N‐enriched and 13 C/ 15 N‐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 1 H, 15 N and 13 C resonances. From both 3D heteronuclear 1 H‐NOE 1 H‐ 13 C and 1 H‐NOE 1 H‐ 15 N 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 Cα atoms is less than 0.12 nm. The secondary structure topology of the molecule is that of an open‐face β sandwich formed by four antiparallel β strands packed against three α 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).