This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Maurer, T.
Right arrow Articles by Kalbitzer, H. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Maurer, T.
Right arrow Articles by Kalbitzer, H. R.

 Previous Article  |  Next Article 

Journal of Bacteriology, September 2004, p. 5906-5918, Vol. 186, No. 17
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.17.5906-5918.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

High-Resolution Structure of the Histidine-Containing Phosphocarrier Protein (HPr) from Staphylococcus aureus and Characterization of Its Interaction with the Bifunctional HPr Kinase/Phosphorylase

Till Maurer,1,{dagger} Sebastian Meier,1,{ddagger} Norman Kachel,1 Claudia Elisabeth Munte,1,§ Sonja Hasenbein,2 Brigitte Koch,2 Wolfgang Hengstenberg,2 and Hans Robert Kalbitzer1*

Institut für Biophysik und Physikalische Biochemie, Universität Regensburg, Regensburg,1 Fakultät für Biologie, Ruhr-Universität Bochum, Bochum, Germany2

Received 5 January 2004/ Accepted 17 May 2004

A high-resolution structure of the histidine-containing phosphocarrier protein (HPr) from Staphylococcus aureus was obtained by heteronuclear multidimensional nuclear magnetic resonance (NMR) spectroscopy on the basis of 1,766 structural restraints. Twenty-three hydrogen bonds in HPr could be directly detected by polarization transfer from the amide nitrogen to the carbonyl carbon involved in the hydrogen bond. Differential line broadening was used to characterize the interaction of HPr with the HPr kinase/phosphorylase (HPrK/P) of Staphylococcus xylosus, which is responsible for phosphorylation-dephosphorylation of the hydroxyl group of the regulatory serine residue at position 46. The dissociation constant Kd was determined to be 0.10 ± 0.02 mM at 303 K from the NMR data, assuming independent binding. The data are consistent with a stoichiometry of 1 HPr molecule per HPrK/P monomer in solution. Using transversal relaxation optimized spectroscopy-heteronuclear single quantum correlation, we mapped the interaction site of the two proteins in the 330-kDa complex. As expected, it covers the region around Ser46 and the small helix b following this residue. In addition, HPrK/P also binds to the second phosphorylation site of HPr at position 15. This interaction may be essential for the recognition of the phosphorylation state of His15 and the phosphorylation-dependent regulation of the kinase/phosphorylase activity. In accordance with this observation, the recently published X-ray structure of the HPr/HPrK core protein complex from Lactobacillus casei shows interactions with the two phosphorylation sites. However, the NMR data also suggest differences for the full-length protein from S. xylosus: there are no indications for an interaction with the residues preceding the regulatory Ser46 residue (Thr41 to Lys45) in the protein of S. xylosus. In contrast, it seems to interact with the C-terminal helix of HPr in solution, an interaction which is not observed for the complex of HPr with the core of HPrK/P of L. casei in crystals.

* Corresponding author. Mailing address: Universität Regensburg, Institut für Biophysik und Physikalische Biochemie, Universitätsstr. 31, D-93040 Regensburg, Germany. Phone: 49 941 943 2594. Fax: 49 941 943 2479. E-mail: hans-robert.kalbitzer{at}biologie.uni-regensburg.de.

{dagger} Present address: Department of Lead Discovery, Boehringer Ingelheim Pharma GmbH & Co. KG, D-88397 Biberach, Germany.

{ddagger} Present address: Biozentrum der Universität Basel, Basel, Switzerland.

§ Present address: Instituto de Física de São Carlos, Universidade de São Paulo, 13560-970 São Carlos SP, Brazil.

Journal of Bacteriology, September 2004, p. 5906-5918, Vol. 186, No. 17
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.17.5906-5918.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kremer, W., Kachel, N., Kuwata, K., Akasaka, K., Kalbitzer, H. R. (2007). Species-specific Differences in the Intermediate States of Human and Syrian Hamster Prion Protein Detected by High Pressure NMR Spectroscopy. J. Biol. Chem. 282: 22689-22698 [Abstract] [Full Text]  
  • Deutscher, J., Francke, C., Postma, P. W. (2006). How Phosphotransferase System-Related Protein Phosphorylation Regulates Carbohydrate Metabolism in Bacteria. Microbiol. Mol. Biol. Rev. 70: 939-1031 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»