Kinetics and thermodynamics of oxygen, CO, and azide binding by the subcomponents of soybean leghemoglobin.
Leghemoglobin shows extreme high affinity behavior in the binding of both oxygen and CO. We have determined the temperature dependence of the rate constants for ligation of oxygen and CO and from these data the thermodynamics (delta G0, delta H0, delta S0) of ligation for the purified components of soybean leghemoglobin. X-ray crystallography has shown that the heme cavity can easily accommodate ligands the size of nicotinate, and analysis of extended x-ray absorption fine structure data has shown that the Fe atom is in the mean plane of the heme in the leghemoglobin-CO complex. Ligation of oxygen and CO are in accord with this picture in that the Ea for oxygen binding is that expected for a diffusion controlled reaction and delta S0 for the ligation of both CO and oxygen is consistent with the simple immobilization of the ligand at the Fe, with no evidence for significant conformational changes in the protein or changes in solvation. At 20 degrees C the rate constants for oxygen and CO binding vary by 26-44% among the eight leghemoglobin components. For azide binding the variation is a factor of 2. These variations appear to arise from amino acid substitutions outside either the heme cavity or the two major paths for ligand entry to the heme. The distribution of leghemoglobin components varies with the age of the soybean nodule during the growing season. The changes in composition alone, however, would only allow the concentration of free oxygen to vary by about 3%. This finding calls into question models that ascribe a significant functional role to changes in the distribution of leghemoglobin components in regulating oxygen concentration in the nodule.
Martin
K D
KD
Department of Chemistry, University of Nebraska, Lincoln 68588-0304.
Saari
L
L
Wang
G X
GX
Wang
T
T
Parkhurst
L J
LJ
Klucas
R V
RV
D001386:Azides / Q000378:metabolism*; D002248:Carbon Monoxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007874:Leghemoglobin / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D019810:Sodium Azide; D013025:Glycine max / Q000032:analysis*; D013696:Temperature; D013816:Thermodynamics
D001386:Azides; D007874:Leghemoglobin; D002248:Carbon Monoxide; D019810:Sodium Azide; D010100:Oxygen