Low-temperature formation of a distal histidine complex in hemoglobin: a probe for heme pocket flexibility.
Pocket dynamics of horse deoxyhemoglobin and methemoglobin in the temperature range from 80 to 260 K is investigated. In both hemoglobins reversible conversion to a low-spin iron complex is observed at temperatures as low as 210 K. Electron spin resonance (ESR) and Mössbauer data assigned this low-spin iron complex to the coordination of N tau-His-E7 as a sixth nitrogenous ligand. The bonding of this ligand located 4 A from the iron indicates the presence of a thermally available conformation that exhibits a high degree of flexibility in the heme pocket. In deoxyhemoglobin, the formation of the bis(histidine) complex was accompanied by excitations of conformational fluctuations manifested through the temperature dependence of the Mössbauer-Lamb factor. The rate for the formation of this complex, with an associated energy barrier (greater than 60 KJ mol-1), is shown to serve as an index of heme pocket flexibility. Measurements performed on partially liganded (carbonmonoxy) hemoglobin indicate that partial ligation enhances conversion of the unliganded subunits to the bis(histidine) complex, suggesting that pocket dynamics is affected by subunit interactions.
D000818:Animals; D001665:Binding Sites; D005615:Freezing; D006418:Heme / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006639:Histidine; D006736:Horses; D008706:Methemoglobin / Q000378:metabolism; D011485:Protein Binding; D013057:Spectrum Analysis
D006454:Hemoglobins; D006418:Heme; D006639:Histidine; D008706:Methemoglobin