Isohemoglobin differentiation in the bimodal-breathing amazon catfish Hoplosternum littorale.
The bimodal gill(water)/gut(air)-breathing Amazonian catfish Hoplosternum littorale that frequents hypoxic habitats uses "mammalian" 2,3-diphosphoglycerate (DPG) in addition to "piscine" ATP and GTP as erythrocytic O(2) affinity modulators. Its electrophoretically distinct anodic and cathodic hemoglobins (Hb(An) and Hb(Ca)) were isolated for functional and molecular characterization. In contrast to Hb(An), phosphate-free Hb(Ca) exhibits a pronounced reverse Bohr effect (increased O(2) affinity with decreasing pH) that is obliterated by ATP, and opposite pH dependences of K(T) (O(2) association constant of low affinity, tense state) and the overall heat of oxygenation. Dose-response curves indicate small chloride effects and pronounced and differentiated phosphate effects, DPG < ATP < GTP < IHP. Hb(Ca)-O(2) equilibria analyzed in terms of the Monod-Wyman-Changeux model show that small T state bond energy differences underlie the differentiated phosphate effects. Synthetic peptides, corresponding to N-terminal fragment of the cytoplasmic domain of trout band 3 protein, undergo oxygenation-linked binding to Hb(Ca), suggesting a metabolic regulatory role for this hemoglobin. The amino acid sequences for the alpha and beta chains of Hb(Ca) obtained by Edman degradation and cDNA sequencing show unusual substitutions at the phosphate-binding site that are discussed in terms of its reverse Bohr effect and anion sensitivities.
Weber
R E
RE
Department of Zoophysiology, University of Aarhus, DK 8000 Aarhus C, Denmark. roy.weber@biology.au.dk
Fago
A
A
Val
A L
AL
Bang
A
A
Van Hauwaert
M L
ML
Dewilde
S
S
Zal
F
F
Moens
L
L
D019794:2,3-Diphosphoglycerate / Q000097:blood; D000595:Amino Acid Sequence; D000811:Anguilla; D000818:Animals; D002397:Catfishes / Q000502:physiology*; D004912:Erythrocytes / Q000502:physiology*; D005880:Gills / Q000502:physiology; D005914:Globins / Q000737:chemistry / Q000378:metabolism*; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010101:Oxygen Consumption; D010108:Oxyhemoglobins / Q000378:metabolism; D012119:Respiration; D012474:Salmon; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D006454:Hemoglobins; D010108:Oxyhemoglobins; D019794:2,3-Diphosphoglycerate; D005914:Globins