Hydrogen ion titrations of the anodic and cathodic haemoglobin components of the European eel Anguilla anguilla.
H+ titrations were conducted on the separated haemoglobin components of eel Anguilla anguilla in both the oxygenated and deoxygenated states. In anodic haemoglobin, the addition of GTP, and to a lesser extent C1-, increased the magnitude of the Haldane effect and shifted its maximum value into the in vivo pH range. Of the 22 histidine residues in the anodic component, only approximately seven were titratable, presumably the beta-chain residues at positions 41, 97, 109 and 146 (helical positions C7, FG4, G11 and HC3, respectively). In cathodic haemoglobin, a small negative Haldane effect was observed at pH values between 6.8 and 8.5 which disappeared in the presence of GTP (molar ratio 3:1 GTP:haemoglobin tetramer). GTP had virtually no effect on the buffer value at fixed oxygenation status, and the lowest buffer value was observed at in vivo pH values. No titratable histidine residues were observed in the cathodic component, indicating that all 14 histidines in this component are buried. We conclude that the anodic component, which constitutes two-thirds of the haemoglobin in the eel, plays the predominant role in CO2 transport and pH homeostasis in vivo.
Brauner
C J
CJ
Danish Center for Respiratory Adaptation, Department of Zoophysiology, Aarhus University, Building 131, Denmark. braunerc@mcmail.cis.mcmaster.ca
Weber
R E
RE
D000811:Anguilla / Q000097:blood*; D000818:Animals; D002021:Buffers; D002852:Chromatography, Ion Exchange; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006454:Hemoglobins / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D010108:Oxyhemoglobins / Q000737:chemistry; D011189:Potassium Chloride / Q000494:pharmacology
D002021:Buffers; D006454:Hemoglobins; D010108:Oxyhemoglobins; D006639:Histidine; D011189:Potassium Chloride; D006160:Guanosine Triphosphate