Relationship between protein/solvent proton exchange and progressive conformation and fluctuation changes in hemoglobin.
The pH dependence of the tertiary structural changes and the quaternary reorganization of the alpha beta interface of oxyhemoglobin in solution has been previously observed in our laboratory. The present work aims to establish whether or not these progressive structural changes with pH result from proton exchange between the protein and the solvent. We therefore have used infrared spectroscopy, acid/base titration and 1H/2H exchange to assess the effect of external proton concentration on the structural and dynamic properties of the hemoglobin molecule in solution. This study is also performed on the carbonmonoxy form since the affinity of hemoglobin for CO is much higher than for oxygen. The present findings demonstrate that affinity changes are closely related to protein fluctuations, as well as structural modifications. An increased affinity, induced either by ligand replacement or pH variation, is associated with the constrained chains exhibiting a lower degree of fluctuation, together with a looser alpha beta interface association.
el Antri
S
S
Laboratoire de Biologie Physico-Chimique, Université Paris VII, France.
Sire
O
O
Alpert
B
B
D003545:Cysteine / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D010108:Oxyhemoglobins / Q000032:analysis; D011487:Protein Conformation; D012997:Solvents; D013055:Spectrophotometry, Infrared
D006454:Hemoglobins; D010108:Oxyhemoglobins; D012997:Solvents; D003545:Cysteine