TY - JOUR
T1 - Allosteric modulation by tertiary structure in mammalian hemoglobins
T2 - Introduction of the functional characteristics of bovine hemoglobin into human hemoglobin by five amino acid substitutions
AU - Fronticelli, Clara
AU - Sanna, Maria Teresa
AU - Perez-Alvarado, Gabriela C.
AU - Karavitis, Michael
AU - Lu, A. Lien
AU - Brinigar, William S.
PY - 1995/12/22
Y1 - 1995/12/22
N2 - Bovine erythrocytes do not contain 2,3-diphosphoglycerate, the principal allosteric effector of human hemoglobin. Bovine hemoglobin has a lower oxygen affinity than human hemoglobin and is regulated by physiological concentrations of chloride (Fronticelli, C., Bucci, E., and Razynska, A. (1988) J. Mol. Biol. 202, 343-348). It has been proposed that the chloride regulation in bovine hemoglobin is introduced by particular amino acid residues located in the amino-terminal region of the A helix and in the E helix of the β subunits (Fronticelli, C. (1990) Biophys. Chem. 37, 141-146). In accordance with this proposal we have constructed two mutant human hemoglobins, β(V1M+H2deleted+T4I+P5A) and β(V1M+H2deleted+T4I+P5A+A76K). These are the residues present at the proposed locations in bovine hemoglobin except for isoleucine at position 4. Oxygen binding studies demonstrate that these mutations have introduced into human hemoglobin the low oxygen affinity and chloride sensitivity of bovine hemoglobin and reveal the presence of a previously unrecognized allosteric mechanism of oxygen affinity regulation where all the interactions responsible for the lowered affinity and chloride binding appear to be confined to individual β subunits.
AB - Bovine erythrocytes do not contain 2,3-diphosphoglycerate, the principal allosteric effector of human hemoglobin. Bovine hemoglobin has a lower oxygen affinity than human hemoglobin and is regulated by physiological concentrations of chloride (Fronticelli, C., Bucci, E., and Razynska, A. (1988) J. Mol. Biol. 202, 343-348). It has been proposed that the chloride regulation in bovine hemoglobin is introduced by particular amino acid residues located in the amino-terminal region of the A helix and in the E helix of the β subunits (Fronticelli, C. (1990) Biophys. Chem. 37, 141-146). In accordance with this proposal we have constructed two mutant human hemoglobins, β(V1M+H2deleted+T4I+P5A) and β(V1M+H2deleted+T4I+P5A+A76K). These are the residues present at the proposed locations in bovine hemoglobin except for isoleucine at position 4. Oxygen binding studies demonstrate that these mutations have introduced into human hemoglobin the low oxygen affinity and chloride sensitivity of bovine hemoglobin and reveal the presence of a previously unrecognized allosteric mechanism of oxygen affinity regulation where all the interactions responsible for the lowered affinity and chloride binding appear to be confined to individual β subunits.
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U2 - 10.1074/jbc.270.51.30588
DO - 10.1074/jbc.270.51.30588
M3 - Article
C2 - 8530494
AN - SCOPUS:0029561342
SN - 0021-9258
VL - 270
SP - 30588
EP - 30592
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 51
ER -