| Popis: |
The dimer-tetramer association constants of several recombinant human hemoglobins (in the CO form) have been measured by differential gel filtration. Recombinant human hemoglobin prepared from recombinant β-chains, and mutant hemoglobins where the substitution was on the surface, β(Thr4 → Asp), in the heme pocket, β(Val67 → Thr), at the 2,3-DPG binding site, β(Val1 → Met + His2del), had a twofold smaller association with respect to natural hemoglobin. In a mutant at the α 1 β 2 1 interface, β(Cys93 → Ala), the association constant was decreased three-fold. Conversely, in a mutant at the α 1 β 1 interface, β(Cys112 → Gly), the association constant was two- and four-fold increased with respect to natural and recombinant human hemoglobin. These differences are energetically very small, consistent with the correct folding of the recombinant hemoglobins. The stabilization of the tetrameric structure by a mutation at the α 1 β 1 interface indicates that structural changes at this interface can be propagated through the protein to the α 1 β 2 interface and, thereby, exert an effect on the allosteric equilibrium. |
