ATP-induced temperature independence of hemoglobin–O2 affinity in heterothermic billfish

Abstract

SUMMARY The inverse relationship between temperature and hemoglobin–O2 affinity resulting from the exothermic nature of heme oxygenation favors O2 unloading from blood to warm, metabolically active tissues. However, this temperature sensitivity is maladaptive, and commonly countered in regional heterotherms, where it may hamper unloading (e.g. in cold extremities of arctic mammals) or increase the diffusive arterio-venous short-circuiting of O2 (e.g. in counter-current heat exchangers of warm swimming muscles of tuna). We hypothesized analogous blood specializations in heterothermic billfish, whose warm eyes and brains increase the temporal resolution of vision, and measured hemoglobin–O2 binding properties in three species over a wide pH range, at two temperatures, and in the absence and presence of the major red cell effector, ATP, permitting detailed assessment of overall oxygenation enthalpies (ΔH′) and contributions from oxygenation-linked proton and ATP dissociation. Billfish express multiple isohemoglobins with similar O2 affinities and pronounced sensitivities to pH and ATP. Compared with the moderate effects associated with proton dissociation upon oxygenation, dissociation of ATP and coupled extra Bohr protons virtually obliterates the temperature sensitivities. At pH 7.4, where this effect is maximal, ATP changes ΔH′ values of blue marlin, striped marlin and shortbill spearfish hemoglobins from −39, −49 and −44 kJ mol−1 O2, respectively, to +26, +4 and −7 kJ mol−1. Thus in addition to allosterically modulating hemoglobin–O2 affinity, ATP diminishes its temperature sensitivity, reducing deleterious arterio-venous short-circuiting of oxygen in the cranial billfish heat exchangers. The mechanism underlying this reduction in oxygenation enthalpy differs fundamentally from that in tuna, supporting independent evolution of this trait in these scombroid lineages.