Oxygen binding and aggregation of hemoglobin from the common European frog, Rana temporaria

Comp Biochem Physiol B Biochem Mol Biol. 1997 Jun;117(2):225-31. doi: 10.1016/s0305-0491(97)00094-1.

Abstract

The hemoglobin from the European frog, Rana temporaria, consists of one major and three minor components. The tetramers aggregate upon deoxygenation notably at pH 7:3. Aggregation due to formation of disulphide bridges, as occurs in related species, was observed only in polyacrylamide gels. The hemolysate showed a pronounced Bohr effect. Oxygen affinity decreased with increasing hemoglobin concentration, indicating that aggregation affects the functional properties of the hemolysate. Oxygen binding equilibria of unfractionated hemolysate are insensitive to chloride and show low sensitivity to ATP. Analysis of oxygen equilibrium curves in terms of the two-state allosteric model (MWC) showed that pH change exerted a greater effect on the association constant of the oxygenated state (KR) than that of the deoxy state (KT). The number of interacting binding sites (q) increased with hemoglobin concentration. Cooperativity of oxygen binding, evaluated as Hill coefficient n, never exceeded the value of 3.0. Earlier studies on hemoglobin and blood from this and related species, report significantly higher n values at high O2 saturation. Molecular adaptive aspects are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Gel
  • Electrophoresis, Polyacrylamide Gel
  • Hemoglobins / chemistry*
  • Hemoglobins / metabolism*
  • Hydrogen-Ion Concentration
  • Isoelectric Focusing
  • Macromolecular Substances
  • Mercaptoethanol / pharmacology
  • Molecular Weight
  • Oxygen / blood*
  • Pulmonary Gas Exchange
  • Rana temporaria / blood*
  • Rana temporaria / physiology

Substances

  • Hemoglobins
  • Macromolecular Substances
  • Mercaptoethanol
  • Oxygen