Autoxidation of giant extracellular hemoglobin of Glossoscolex paulistus: molecular mechanism and oligomeric implications.

Giant extracellular hemoglobins present high redox stability due to their supramolecular architecture, high number of polypeptide chains and great compaction of protein subunits. The oligomeric assembly and the changes in the polypeptidic structure can influence the autoxidation rate of the heme proteins, being that different nucleophiles can act in this process due to pH alterations. In the present work, we have studied the autoxidation rate of whole Glossoscolex paulistus (HbGp) giant extracellular hemoglobin, as well as the autoxidation rate of the isolated d monomer of HbGp studied regarding pH variations.

Ferric species of the giant extracellular hemoglobin of Glossoscolex paulistus as function of pH: an EPR study on the irreversibility of the heme transitions.

The present article is focused on the transitions of ferric heme species of the giant extracellular hemoglobin of Glossoscolex paulistus (HbGp) induced by successive alterations in pH, involving alkaline and acid mediums. Electron paramagnetic resonance (EPR) is the spectroscopy used to evaluate the transitions that occur in the first coordination sphere of ferric ion as a consequence of ligand changes in a wide range of pH, since this tool is very sensitive to slight changes that occur in the heme pocket of paramagnetic species. This approach is adequate to obtain information regarding the reversibility/irreversibility that involves the heme transitions induced by pH, since the degree of reversibility is associated to the intensity of the changes that occur in the spatial configuration of the polypeptide chains, which is clearly associated to the first coordination sphere.

Ferric species equilibrium of the giant extracellular hemoglobin of Glossoscolex paulistus in alkaline medium: HALS hemichrome as a precursor of pentacoordinate species.

The present work is focused on the complex ferric heme species equilibrium of the giant extracellular hemoglobin from Glossoscolex paulistus (HbGp) in alkaline medium. EPR, UV-vis and CD spectroscopies were used in order to characterize the ferric heme species formed as a consequence of the medium alkalization as well as the oligomeric changes occurring simultaneously with heme transitions. EPR experiments allowed us to characterize the different hemichrome species in equilibrium, illustrating the small difference in spin state of this species and the complexity of the equilibira involving hemoglobin ferric species.

Pentacoordinate and hexacoordinate ferric hemes in acid medium: EPR, UV-Vis and CD studies of the giant extracellular hemoglobin of Glossoscolex paulistus.

The equilibrium complexity involving different axially coordinated hemes is peculiar to hemoglobins. The pH dependence of the spontaneous exchange of ligands in the extracellular hemoglobin from Glossoscolex paulistus was studied using UV-Vis, EPR, and CD spectroscopies. This protein has a complex oligomeric assembly with molecular weight of 3.

Autoxidation studies of extracellular hemoglobin of Glossoscolex paulistus at pH 9: cyanide and hydroxyl effect.

The complex oligomeric assembly of the hemoglobin subunits may influence the autoxidation rate. To understand this relation, the rate of autoxidation was studied at pH 9.0, where the Glossoscolex paulistus Hemoglobin (GpHb) dissociates.