title: Alteration of hemoglobin function by diadenosine 5',5'''-P1,P4-tetraphosphate and other alarmones.
abstract: Heat-shocked organisms are known to produce not only "heat shock proteins" but also diadenosine tetraphosphate (Ap4A) and related compounds that may act as "alarmones" that alert the cell to the onset of metabolic stress. We found that Ap4A is synthesized in chicken erythrocytes and that the Ap4A level in the whole blood of heat-stressed birds increases about 10-fold. In searching for alarmone receptors, we found that the diadenosine polyphosphates bind preferentially with high affinity to the deoxy conformation of hemoglobin in a ratio of one/tetramer. The binding affinity of this new class of effectors of hemoglobin function is directly related to the number of phosphates which bridge the nucleotide moieties, with the most dramatic in vitro effect on oxygen affinity being shown by Ap6A. Decreasing effects are brought about by diadenosine penta-, tetra-, tri-, di-, and monophosphates. The association constant for Ap4A binding to deoxygenated human hemoglobin at pH 7.25 is 26 microM-1, close to that for 2,3-diphosphoglycerate. At 100-fold excess over heme, Ap4A increases the P50 of stripped Hb A in 0.05 M HEPES buffer at pH 7.25, 20 degrees C, from 0.85 to 6.03 mm Hg. The binding, which markedly enhances the Bohr effect, involves the beta chain anion-binding site. The kinetics of both ligand binding and dissociation are affected, with a greater quantitative effect on the oxygen dissociation process. Although the low concentration of the diadenosine polyphosphates in red cells precludes a physiologically significant modulation of oxygen delivery, competition with the ATP- and NAD(P)H-binding sites on hemoglobin or regulatory enzymes may prove to be of adaptive significance.
authors.affiliation: Marine Biomedical Center, Duke University Marine Laboratory, Beaufort, North Carolina 28516.
authors.forename: C
authors.initials: C
authors.lastname: Bonaventura
authors.forename: R
authors.initials: R
authors.lastname: Cashon
authors.forename: J M
authors.initials: JM
authors.lastname: Colacino
authors.forename: R H
authors.initials: RH
authors.lastname: Hilderman
mesh_terms: D000818:Animals; D001717:Birds; D002248:Carbon Monoxide / Q000378:metabolism; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008968:Molecular Conformation; D010100:Oxygen / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D015226:Dinucleoside Phosphates; D006454:Hemoglobins; C020733:diadenosine tetraphosphate; D002248:Carbon Monoxide; D010100:Oxygen