Characterization of an artificial dimer of ribonuclease H using 1H NMR spectroscopy

J Biomol NMR. 1996 Jan;7(1):29-34. doi: 10.1007/BF00190454.

Abstract

The protein fusion technique was applied in the synthesis of an artificial dimer of ribonuclease H (305 residues). 1H NMR spectroscopy was used to analyze the structure of this dimer. Spectral profiles and pKa values of the histidine residues obtained using 1H NMR indicate that the dimer retains the secondary and tertiary structures of the intact monomer. Selective spin-lattice relaxation measurements suggest that the two monomeric units in the dimer are in tight contact. Furthermore, the 2D 1H NMR and paramagnetic relaxation filter results show that the two monomers bind together through interactions between the N- and C-terminal sites of the linked regions.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Cloning, Molecular
  • Histidine
  • Hydrogen
  • Hydrogen-Ion Concentration
  • Kinetics
  • Macromolecular Substances
  • Magnetic Resonance Spectroscopy
  • Peptide Fragments / chemistry
  • Plasmids
  • Protein Conformation*
  • Recombinant Proteins / chemistry
  • Ribonuclease H / chemistry*
  • Sequence Deletion

Substances

  • Macromolecular Substances
  • Peptide Fragments
  • Recombinant Proteins
  • Histidine
  • Hydrogen
  • Ribonuclease H