pH dependence of the dissociation of multimeric hemoglobin probed by high hydrostatic pressure.
We investigated the thermodynamic features of the classic alkaline dissociation of multimeric hemoglobin (3.1 MDa) from Glossoscolex paulistus (Annelidea) using high hydrostatic pressure. Light scattering measurements up to microscopic thermodynamic equilibrium indicated a high pH dependency of dissociation and association. Electron microscopy and gel filtration corroborated these findings. The volume change of dissociation decreased in absolute values from -48.0 mL/mol of subunit at pH 6.0 to -19.2 mL/mol at pH 9.0, suggesting a lack of protein interactions under alkaline conditions. Concomitantly, an increase in pH reduced the Gibbs free energy of dissociation from 37.7 to 27.5 kJ/mol of subunit. The stoichiometry of proton release calculated from the pressure-induced dissociation curves was +0.602 mol of H(+)/mol of subunit. These results provide a direct quantification of proton participation in stabilizing the aggregated state of the hemoglobin, and contribute to our understanding of protein-protein interactions and of the surrounding conditions that modulate the process of aggregation.
Bispo
Jose A C
JA
Laboratório de Termodinâmica de Proteínas, Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, Campinas, SP, CEP 13083-970, Brazil.
Santos
Jose L R
JL
Landini
Gustavo F
GF
Goncalves
Juliana M
JM
Bonafe
Carlos F S
CF
D019281:Dimerization; D006454:Hemoglobins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006874:Hydrostatic Pressure; D013816:Thermodynamics
D006454:Hemoglobins