Open AccessThree‐dimensional 15 N‐ 1 H‐ 1 H and 15 N‐ 13 C‐ 1 H nuclear‐magnetic resonance studies of HPr a central component of the phospho enol pyruvate‐dependent phosphotransferase system from Escherichia coli
Author(s) -
NULAND Nico A. J.,
DIJK Alard A.,
DIJKSTRA Klaas,
HOESEL Frans H. J.,
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.tb16573.x
Subject(s) - heteronuclear molecule , chemistry , spectroscopy , nuclear magnetic resonance spectroscopy , carbon 13 , two dimensional nuclear magnetic resonance spectroscopy , nuclear magnetic resonance , antiparallel (mathematics) , crystallography , geminal , analytical chemistry (journal) , stereochemistry , physics , quantum mechanics , magnetic field , chromatography
We have performed three‐dimensional NMR studies on a central component of the phospho enol pyruvate‐dependent phosphotransferase system of Escherichia coli , denoted as HPr. The protein was uniformly enriched with 15 N and 13 C to overcome spectral overlap. Complete assignments were obtained for the backbone 1 H, 15 N and 13 C resonances, using three‐dimensional heteronuclear 1 H NOE 1 H‐ 15 N multiple‐quantum coherence spectroscopy (3D‐NOESY‐HMQC) and three‐dimensional heteronuclear total correlation 1 H‐ 15 N multiple‐quantum coherence spectroscopy (3D‐TOCSY‐HMQC) experiments on 15 N‐enriched HPr and an additional three‐dimensional triple‐resonance 1 HN‐ 15 N‐ 13 Cα correlation spectroscopy (HNCA) experiment on 13 C, 15 N‐enriched HPr. Many of the sequential backbone 1 H assignments, as derived from two‐dimensional NMR studies [Klevit, R. E., Drobny, G. P. & Waygood, E. B. (1986) Biochemistry 25 , 7760–7769], were corrected. Almost all discrepancies are in the helical regions, leaving the published antiparallel β‐sheet topology almost completely intact.