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The structure and function of HPr

E B Waygood1

  • 1Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada. waygood@sask.usask.ca

Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire
|January 29, 1999
PubMed
Summary

Histidine-containing phosphocarrier protein (HPr) structure is conserved across species, revealed by NMR. Phosphorylation alters the active site conformation of HPr, impacting its function.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Nuclear Magnetic Resonance (NMR) Spectroscopy

Background:

  • Histidine-containing phosphocarrier protein (HPr) is crucial in bacterial phosphotransferase systems.
  • Early 2D 1H-NMR studies elucidated HPr's tertiary structure, revealing a conserved betaalpha betabeta alphabeta alpha fold.

Purpose of the Study:

  • To detail the high-resolution structures of HPr from Escherichia coli and Bacillus subtilis using isotopic labeling.
  • To characterize the active site of HPr and its phosphorylated form (phospho-HPr), including unusual pKa values.

Main Methods:

  • Two-dimensional 1H-NMR spectroscopy was employed to determine HPr tertiary structures.
  • 15N- and 13C-labeled proteins were utilized for high-resolution structural analysis.
  • NMR was applied to identify the phosphohistidine isomer and analyze pKa values of His-15.

Main Results:

  • The overall folding pattern of HPr, comprising alpha-helices and a beta-sheet, is highly conserved across different bacterial species.
  • NMR revealed the Ndelta1-P-histidine isomer in S. aureus phospho-HPr and unusual pKa values for His-15.
  • His-15 in HPr is conformationally stable, affected by helix A's dipole, and undergoes a slight shift upon phosphorylation to interact with specific residues and the phosphoryl group.

Conclusions:

  • The conserved structure of HPr suggests a fundamental role in phosphotransferase systems.
  • Phosphorylation induces subtle yet significant conformational changes in the HPr active site, influencing its interactions and function.
  • NMR spectroscopy is a powerful tool for elucidating protein structure, dynamics, and the impact of post-translational modifications like phosphorylation.

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