Characterization of the hemoglobins of the Australian lungfish Neoceratodus forsteri (Krefft).
We examined for the first time the hemoglobin components of the blood of the Australian lungfish, Neoceratodus forsteri and their functional responses to pH and the allosteric modulators adenosine triphosphate (ATP), guanosine triphosphate (GTP), 2,3-bisphosphoglyceric acid (BPG) and inositol hexaphosphate (IHP) at 25 degrees C. Lysates prepared from stripped, unfractionated hemolysate produced sigmoidal oxygen equilibrium curves with high oxygen affinity (oxygen partial pressure required for 50% hemoglobin saturation, p(50)=5.3 mmHg) and a Hill coefficient of 1.9 at pH 7.5. p(50) was 8.3 and 4.5 mmHg at pH 6 and 8, respectively, which corresponded to a modest Bohr coefficient (Delta log p(50)/Delta pH) of -0.13. GTP increased the pH sensitivity of oxygen binding more than ATP, such that the Bohr coefficient was -0.77 in the presence of 2 mmol L(-1) GTP. GTP was the most potent regulator of hemoglobin affinity, with concentrations of 5 mmol L(-1) causing an increase in p(50) from 5 to 19 mm Hg at pH 7.5, while the order of potency of the other phosphates was IHP>ATP>BPG. Three hemoglobin isoforms were present and each contained both alpha and beta chains with distinct molecular weights. Oxygen affinity and pH-dependence of isoforms I and II were essentially identical, while isoform III had a lower affinity and increased pH-dependence. The functional properties of the hemoglobin system of Neoceratodus appeared consistent with an active aquatic breather adapted for periodic hypoxic episodes.
Department of Zoophysiology, Institute of Biological Sciences, University of Aarhus, Aarhus, DK8000, Denmark.
Jonas R
JR
Rasmussen
Rufus M G
RM
Wells
Kristen
K
Henty
Timothy D
TD
Clark
Thomas
T
Brittain
D019794:2,3-Diphosphoglycerate / Q000097:blood; D000222:Adaptation, Physiological; D000255:Adenosine Triphosphate / Q000097:blood; D000818:Animals; D005399:Fishes / Q000097:blood* / Q000502:physiology; D006160:Guanosine Triphosphate / Q000097:blood; D006454:Hemoglobins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D010100:Oxygen / Q000097:blood*; D010108:Oxyhemoglobins / Q000737:chemistry / Q000378:metabolism*; D010313:Partial Pressure; D010833:Phytic Acid / Q000097:blood; D011487:Protein Conformation; D020033:Protein Isoforms; D012965:Sodium Chloride / Q000097:blood; D013329:Structure-Activity Relationship
D006454:Hemoglobins; D010108:Oxyhemoglobins; D020033:Protein Isoforms; D019794:2,3-Diphosphoglycerate; D012965:Sodium Chloride; D010833:Phytic Acid; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010100:Oxygen