Oxygen binding and its allosteric control in hemoglobin of the pulmonate snail, Biomphalaria glabrata.

Abstract

Pulmonate snails that experience extreme variations in gas tensions and temperatures possess extracellular, high-molecular mass ( approximately 1.7 x 10(6) Da) hemoglobins (Hbs) that are little known as regards oxygenation and allosteric characteristics. Biomphalaria glabrata hemolymph exhibits a high O2 affinity (half-saturation O2 tension = 6.1 mmHg; pH 7.7, 25 degreesC), pronounced Bohr effect (Bohr factor = -0.5), and pH-dependent cooperativity (Hill's cooperativity coefficient at half-saturation = 1.1-2.0). Divalent cations increase O2 affinity, Ca2+ exerting greater effect than Mg2+. Analyses in terms of the Monod-Wyman-Changeux model indicate novel O2 affinity control mechanisms. In contrast to vertebrate Hb, where organic phosphates and protons lower affinity via decreased O2 association equilibrium constant of Hb in low-affinity state (KT), and to extracellular annelid Hbs, where protons and cations primarily modulate O2 association equilibrium constant of Hb in high-affinity state (KR), in B. glabrata Hb, the Bohr effect is mediated predominantly via KR and the cation effect via KT, reflecting preferential, oxygenation-linked proton binding to oxygenated Hb and cation binding to deoxygenated Hb. CO2 has no specific (pH independent) effect. Nonlinear van't Hoff plots show temperature dependence of the overall heats of oxygenation, indicating oxy-deoxy heat capacity differences. The findings are related to possible physiological significance in pond habitats.