Solution structure of the histidine-containing phosphocarrier protein fromStaphylococcus carnosus

Abstract

The solution structure of histidine-containing phosphocarrier protein from Staphylococcus carnosus was determined by two-and three-dimensional nuclear magnetic resonance (NMR) spectroscopy on uniformly 15 N-enriched protein. The main structural element is an antiparallel (3-pleated sheet with four strands A, B, C, and D arranged with the topology A-D-B-C. Strand A comprises residues 2 to 8, strand B residues 32 to 37, strand C reidues 40 to 43, and strand D residues 59 to 66. Three right-handed helices are arranged on top of the (3-pleated sheet. Helix a reaches from residue 16 to 29, helix b from residue 48 to 53, and helix c from residue 72 to 83. Strands B and C of the (3-pleated sheet are connected by a type II' turn. The hydroxyl proton of Ser-31 is exchanging with the solvent so slowly that cross peaks can be detected in two-dimensional NMR spectra based on homonuclear J-couplings. The imidazole ring of the active-center His-15, which is partly charged in the structure determined at pH 7.2, is located above the N-terminal end of helix a, perpendicular to its axis. The N5 ' atom of His-15, accepting the phosphoryl from enzyme 1, is exposed to the solvent.

High-Resolution Structure of the Histidine-Containing Phosphocarrier Protein (HPr) fromStaphylococcus aureusand Characterization of Its Interaction with the Bifunctional HPr Kinase/Phosphorylase

“…As described earlier (28), for the HPr protein of S. aureus there is an interesting exception: the cross peaks between the H ␥ proton of the hydroxyl group and the H ␣ and H ␤ protons of Ser31 can be observed directly in the TOCSY spectra at 303 K. This is a typical spectroscopic feature of HPr proteins which has been described for HPr from Staphylococcus carnosus (14) as well as HPr from E. coli (20), which both contain a conserved serine residue at this position. The hydroxyl proton resonances of Ser31 could be observed at 5.58 ppm in HPr from S. carnosus and at 5.77 ppm in HPr from E. coli, very close to the resonance position of 5.56 ppm obtained for HPr from S. aureus.…”

Molecular dynamics simulations of HPr under hydrostatic pressure

“…4 The MD and molecular mechanics calculation were performed with the parm98 parameter set 25 and the TIP3P model for water. 26 The program AMBER 6.0 27 was used for the simulations performed with the Berendsen thermostat, and the program ORAC 4.0 28 for the simulations performed with the NoseHoover thermostat.…”

“…The RMSD values of atomic coordinates between the mean conformers are in the ranges of 0.68 -1.0 Å for all atoms and of 0.57-0.95 Å for heavy atoms: this is of the same order of value as the RMSD values between the conformers of the NMR structure. 4 The global changes induced on the protein structure by the hydrostatic pressure are on average of similar amplitude than the conformational variability in the NMR structure determined at 0.1 MPa. Mean RMSD values of 0.50, 0.64, and 0.63 Å are found for the 2-7, 20 -41, and 50 -84 regions, which are the best defined regions in the NMR structure.…”

“…4 The ␣-helices are colored in red, the ␤-sheet in green, and the loops in light blue. The figure was realized using Molscript v2.1 49 and Raster3D 2.7b.…”

“…The deformations induced on the protein structure at high hydrostatic pressure values (4,50, 100, 150, and 200 MPa) were previously (H. Kalbitzer, A. Görler, H. Li, P. Dubovskii, A. Hengstenberg, C. Kowolik, H. Yamada, and K. Akasaka, Protein Science 2000, Vol. 9, pp.…”