POLAR BEAR HEMOGLOBIN AND HUMAN Hb A₀: SAME 2,3-DIPHOSPHOGLYCERATE BINDING SITE BUT ASYMMETRY OF THE BINDING?

Abstract

Polar bear (Ursus maritimus) hemoglobin (Hb) shows a low response to 2,3-diphosphoglycerate (2,3-DPG), compared to human Hb A₀, even though these proteins have the same 2,3-DPG-binding site. In addition, polar bear Hb shows a high response to chloride and an alkaline Bohr effect (Δlog P₅₀/ΔpH) that is significantly greater than that of human Hb A₀. The difference in sequence Pro (Hb A₀)→Gly (polar bear Hb) at position A2 in the A helix seems to be critical for reduced binding of 2,3-DPG. Our results also show that the A2 position may influence not only the flexibility of the A helix, but that differences in flexibility of the first turn of the A helix may affect the unloading of oxygen for the intrinsic ligand affinities of the α and β chains. However, preferential binding to either chain can only take place if there is appreciable asymmetric binding of the phosphoric effector. Regarding this point, ³¹P NMR data suggest a loss of symmetry of the 2,3-DPG-binding site in the deoxyHb–2,3-DPG complex.