Functional and structural characterization of the myoglobin from the polychaete Ophelia bicornis

Biochem J. 2005 Jul 15;389(Pt 2):497-505. doi: 10.1042/BJ20050267.

Abstract

The myoglobin of the polychaete annelid Ophelia bicornis was isolated, purified to homogeneity and characterized. The primary structure, obtained from cDNA and protein sequencing, consists of 139 amino acid residues. The alignment with other globin sequences showed that O. bicornis myoglobin misses the pre-A helix and the first six residues of the A helix. The presence of a PheB10-GlnE7 haem distal residue pair is in agreement with the measured oxygen affinity (P50=0.85 mmHg; 1 mmHg=0.133 kPa) and the only slightly higher autoxidation rate constant (0.28 h(-1)) with respect to that of the sperm whale myoglobin mutant E7 His-->Gln (0.21 h(-1)) and to elephant myoglobin (0.1 h(-1)). Oxygen-binding co-operativity was found to be absent under all the examined experimental conditions. The resistance of O. bicornis myoglobin towards autoxidation seems to confirm the important role of part of the A helix in the stability of the globin. The higher pKa of the acid-alkaline ferric transition of O. bicornis with respect to Asian elephant myoglobin, as well as the higher absorbance ratio of its ferric form to the oxy form measured in the Soret region (gammamet/gammaoxy) with respect to that of the African elephant myoglobin, suggested a stronger interaction between the distal glutamine and the water molecule at the sixth co-ordinate position.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Regulation
  • Allosteric Site
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • DNA, Complementary / genetics
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Sequence Data
  • Myoglobin / chemistry*
  • Myoglobin / genetics
  • Myoglobin / isolation & purification
  • Myoglobin / metabolism*
  • Oxygen / metabolism
  • Polychaeta / chemistry*
  • Protein Binding
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Electrospray Ionization
  • Titrimetry

Substances

  • DNA, Complementary
  • Myoglobin
  • Oxygen