Deoxygenation-linked association of a tetrameric component of chicken hemoglobin.
Deoxygenation-dependent association of hemoglobin tetramers appears to be widespread among amphibians, reptiles, and possibly all or most birds. The evidence for this conclusion depends largely on oxygen equilibria of whole blood which have Hill coefficients that reach values as high as 5-7 at 80-90% oxygenation. Computer simulation of the sedimentation velocity behavior of the major components A and D of chicken hemoglobin shows that component D but not A self-associates to form dimers of tetramers. The gradient profiles at pH 7.5 were satisfactorily fitted with an association constant of 1.26 x 10(4) M-1 and sedimentation coefficients of 4.63 and 7.35 S for tetramer and (tetramer)2, respectively. Since components A and D share common beta chains we conclude that tetramer-tetramer contacts must depend on surface residues of the alpha chains. Comparison of the amino acid sequences of the alpha D and alpha A chains of the hemoglobins from 12 avian species ranging from sparrow to ostrich shows that 20 residues are conserved in the alpha D chains but not in the alpha A chains. Nine of these (45%) are clustered between positions E20 and FG2. Four of the latter, Lys71 (E20), Asn75 (EF4), Gln78 (EF7), and Glu82 (F3) are conserved in all alpha D chains even though they do not appear to participate in intratetramer contacts. Molecular modeling indicates that residues Lys71, Gln78, and Glu82 of the alpha chain are strong candidates for the primary tetramer-tetramer contacts.
Cobb
J A
JA
Department of Zoology, University of Texas, Austin 78712.
Manning
D
D
Kolatkar
P R
PR
Cox
D J
DJ
Riggs
A F
AF
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002645:Chickens; D002852:Chromatography, Ion Exchange; D006454:Hemoglobins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000378:metabolism*; D011487:Protein Conformation; D014461:Ultracentrifugation
D000596:Amino Acids; D006454:Hemoglobins; C100003:deoxyhemoglobin; D010100:Oxygen