Haemoglobin-Oxygen Binding Properties in the Blood of Xenopus Laevis, with Special Reference to the Influences of Aestivation and of Temperature and Salinity Acclimation

1980 ◽  
Vol 86 (1) ◽  
pp. 19-37
Author(s):  
ALFRED JOKUMSEN ◽  
ROY E. WEBER
Keyword(s):  
Xenopus Laevis ◽  
Salt Water ◽  
Oxygen Affinity ◽  
Minor Component ◽  
Blood Levels ◽  
Oxygen Binding ◽  
Binding Properties ◽  
Salinity Acclimation ◽  
High O2 ◽  
O2 Binding

The oxygen equilibrium properties of blood and of solutions of haemoglobin from Xenopus laevis are reported. At pH 7.6 the oxygen affinity of the blood, expressed as half saturation oxygen tension (P50) amounts to 27.0 mm and 13.7 mmHg (3.60 and 1.83 kPa) when measured at 25 and 10 °C, while the Bohr factor (Δlog P50/ΔpH) was −0.40, and the Hill's cooperativity coefficient, n, averaged 2.1. These data reflect an overall heat of oxygenation, ΔH, of −7.9 kcal. mol−1, which decreased to −6.3 kcal. mol−1 when the live animals were acclimated to each measuring temperature. Xenopus blood showed a high O2 capacity (15 vol.%) compared to that of other amphibians. Acclimation to water of increased salinity (12%0), and aestivation, raised blood O2 affinity; at 25 °C and pH 7.6, P50 decreased to 21.1 and 25.2 mmHg (2.81 and 3.36 kPa), respectively. These changes were concomitant with increases in the blood levels of urea. In contrast to NaCl and ATP, urea increased O2 affinity of the purified haemoglobin, suggesting that oxygenationlinked binding to haemoglobin is involved in the modulations of the blood O2 affinity during aestivation and acclimation to salt water. Xenopus haemoglobin consists of two components. The major component is electrophoretically anodal, and has O2 binding properties similar to those of the haemolysate; the minor component is cathodal, and shows extremely low P50, pH sensitivity and cooperativity.

scholarly journals Haemoglobin polymorphisms affect the oxygen-binding properties in Atlantic cod populations

2008 ◽  
Vol 276 (1658) ◽  
pp. 833-841 ◽  
Author(s):  
Øivind Andersen ◽  
Ola Frang Wetten ◽  
Maria Cristina De Rosa ◽  
Carl Andre ◽  
Cristiana Carelli Alinovi ◽  
...  
Keyword(s):  
Evolutionary Biology ◽  
Quaternary Structure ◽  
Atlantic Cod ◽  
Three Dimensional ◽  
Oxygen Affinity ◽  
Oxygen Binding ◽  
Low Oxygen ◽  
Binding Properties ◽  
Important Species ◽  
The North

A major challenge in evolutionary biology is to identify the genes underlying adaptation. The oxygen-transporting haemoglobins directly link external conditions with metabolic needs and therefore represent a unique system for studying environmental effects on molecular evolution. We have discovered two haemoglobin polymorphisms in Atlantic cod populations inhabiting varying temperature and oxygen regimes in the North Atlantic. Three-dimensional modelling of the tetrameric haemoglobin structure demonstrated that the two amino acid replacements Met55β 1 Val and Lys62β 1 Ala are located at crucial positions of the α 1 β 1 subunit interface and haem pocket, respectively. The replacements are proposed to affect the oxygen-binding properties by modifying the haemoglobin quaternary structure and electrostatic feature. Intriguingly, the same molecular mechanism for facilitating oxygen binding is found in avian species adapted to high altitudes, illustrating convergent evolution in water- and air-breathing vertebrates to reduction in environmental oxygen availability. Cod populations inhabiting the cold Arctic waters and the low-oxygen Baltic Sea seem well adapted to these conditions by possessing the high oxygen affinity Val55–Ala62 haplotype, while the temperature-insensitive Met55–Lys62 haplotype predominates in the southern populations. The distinct distributions of the functionally different haemoglobin variants indicate that the present biogeography of this ecologically and economically important species might be seriously affected by global warming.

Regulation of Blood Oxygen Affinity in the Australian Blackfish Gadopsis Marmoratus: I. Correlations between Oxygen-binding Properties, Habitat and Swimming Behaviour

1982 ◽  
Vol 99 (1) ◽  
pp. 223-243
Author(s):  
G. P. DOBSON ◽  
J. BALDWIN
Keyword(s):  
Whole Blood ◽  
Oxygen Affinity ◽  
Swimming Behaviour ◽  
Oxygen Binding ◽  
Blood Oxygen ◽  
Low Oxygen ◽  
Binding Properties ◽  
Molar Ratios ◽  
Blood Oxygen Affinity ◽  
Low Oxygen Affinity

1. The regulation of whole blood oxygen affinity in the freshwater blackfish Gadopsis marmoratus Richardson has been examined, and correlations made between oxygen-binding properties and the habitat and swimming behaviour of the fish. 2. Blackfish whole blood has a low oxygen affinity relative to other fish bloods reported in the literature. This is not due to a low oxygen affinity of the stripped haemoglobins, but arises from interactions between haemoglobin and intraerythrocytic modulators. 3. The presence of high concentrations of ATP, and to a lesser extent GTP, in the erythrocyte, together with the effect of these nucleoside triphosphates on the oxygen affinity of haemoglobin solutions at physiological NTP: Hb4 molar ratios, demonstrates that this class of compounds is a major regulator of oxygen affinity in blackfish blood. 4. The oxygen affinities of whole blood and haemoglobin solutions are sensitive to pH, with haemoglobin solutions displaying a relatively large alkaline Bohr coefficient of - 1.05 over the physiologically relevant pH range of 6.5–7.0. 5. Although increasing Pco2, lowers the oxygen affinity of whole blood, it does so only through the effect on pH, as pH-buffered haemoglobin solutions show no oxygen-linked CO2 binding. This lack of oxygen-linked CO2 binding has not been reported for any other naturally occurring vertebrate haemoglobins. 6. Muscle morphology and biochemistry, and behavioural observations, indicate that the blackfish uses anaerobic energy metabolism during rapid swimming and in recovery. 7. It is concluded that the oxygen-binding properties of blackfish blood reflect adaptations for maintaining adequate tissue oxygenation for animals at rest and during slow sustained swimming in waters of high oxygen tensions.

scholarly journals Amino Acids Responsible for Decreased 2, 3-Biphosphosphoglycerate Binding to Fetal Hemoglobin

Blood ◽  
1997 ◽  
Vol 90 (8) ◽  
pp. 2916-2920 ◽  
Author(s):  
Kazuhiko Adachi ◽  
Patrick Konitzer ◽  
Jian Pang ◽  
Konda S. Reddy ◽  
Saul Surrey
Keyword(s):  
Expression System ◽  
Oxygen Affinity ◽  
Fetal Hemoglobin ◽  
Oxygen Binding ◽  
Cell Disease ◽  
Binding Properties ◽  
Yeast Expression System ◽  
Normal Human ◽  
Potential Use

Abstract To clarify the role of γN-terminal Gly, γ5 Glu, and γ143 Ser in 2, 3-biphosphosphoglycerate (BPG) binding to fetal hemoglobin (Hb F ), we engineered and produced normal human Hb F and two Hb F variants (Hb F γG1V, γS143H, and Hb F γG1V, γE5P, γS143H) using a yeast expression system and then compared their oxygen-binding properties with those of native human Hb F and adult Hb (Hb A). Oxygen affinity of Hb F γG1V, γS143H in the absence of 2, 3-BPG was slightly higher than that of normal Hb F. The decrease in oxygen affinities for Hb F γG1V, γS143H with increasing 2, 3-BPG concentrations was larger than that of normal Hb F, but significantly less than that of Hb A. In contrast, oxygen affinities of Hb F γG1V, γE5P, γS143H in the absence and presence of 2, 3-BPG were much lower than those of Hb F γG1V, γS143H and were similar to those of Hb A. These results indicate that differences between Pro and Glu at the A2 position in the A helix in Hb A and Hb F, respectively, are critical for reduced binding of 2, 3-BPG to Hb F, even though β5 Pro does not interact directly with 2, 3-BPG in Hb A. Hb F variants such as Hb F γG1V, γE5P, γS143H, which exhibit reduced oxygen affinity, should facilitate design of efficient antisickling fetal Hb variants for potential use in gene therapy for sickle cell disease.

Temperature acclimation and oxygen-binding properties of blood and multiple haemoglobins of rainbow trout

1976 ◽  
Vol 65 (2) ◽  
pp. 333-345 ◽  
Author(s):  
R. E. Weber ◽  
S. C. Wood ◽  
J. P. Lomholt
Keyword(s):  
Rainbow Trout ◽  
Whole Blood ◽  
Relative Concentration ◽  
Oxygen Affinity ◽  
Temperature Acclimation ◽  
Organic Phosphate ◽  
Red Cells ◽  
Oxygen Binding ◽  
Binding Properties

Acclimation of rainbow trout to 5, 15 and 22 degrees C for periods exceeding 4 months had no significant effect on the oxygen affinity of whole blood or on the concentration of ATP, which is the main organic phosphate in red cells. Slight differences were, however, found in the oxygenation properties of the haemolysates, which correlate with changes in the relative concentration of the multiple haemoglobins. The oxygen-binding properties of the main haemoglobin components account for the observed differences in the haemolysates. The possible thermoacclimatory significance of changes in haemoglobin multiplicity and co-factor concentrations is discussed.

scholarly journals The quarternary structure of an unusual high-molecular-weight intracellular haemoglobin from the bivalve mollusc Barbatia reeveana

1980 ◽  
Vol 189 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Nicholas P. Grinich ◽  
Robert C. Terwilliger
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Sodium Dodecyl ◽  
Oxygen Affinity ◽  
Minor Component ◽  
Subunit Structure ◽  
Oxygen Binding ◽  
Low Oxygen ◽  
Unusual Structure ◽  
Binding Domains ◽  
Covalently Linked

The arcid clam Barbatia reeveana contains an intracellular haemoglobin with an unusual structure. First, compared with other intracellular haemoglobins, it is extremely large, with a mol.wt. of 430000 and an s20,w of 13.6S. A minor component (mol.wt.=220000; s20,w=9.7S) is also present as a probable dissociation product of the major component. Secondly, this haemoglobin has an unusual subunit structure. It contains 1mol of haem per 16000g of protein, in common with most other haemoglobins. However, the smallest polypeptide that could be obtained after treatment with sodium dodecyl sulphate or 6m-guanidine with reducing agent has a mol.wt. of 32000–37000. Digestion of the haemoglobin with the proteinase subtilisin produces both 57000- and 30000-mol.wt. aggregates that contain 1mol of haem per 16000g of protein and that can be dissociated into 16500-mol.wt. polypeptides by treatment with sodium dodecyl sulphate. The intact polymer shows slight co-operativity (h=1.7), lacks a Bohr effect between pH7 and 8, and has a low oxygen affinity [P50=4.8kPa (36mmHg) at 20°C] relative to other haemoglobins. The 30000-mol.wt. aggregate obtained by digestion of the polymer binds oxygen reversibly with an affinity greater than that of the polymer, but with some co-operativity (h=1.7). These results are consistent with the hypothesis that the subunits of this unusually large intracellular haemoglobin are 32000-mol.wt. polypeptides that in turn are composed of two covalently linked haem-containing oxygen-binding domains. This is the first report of an intracellular haemoglobin with such a structure.

Oxygen binding by single red blood cells from the red-eared turtle Trachemys scripta

2001 ◽  
Vol 90 (5) ◽  
pp. 1679-1684 ◽  
Author(s):  
Sebastian Frische ◽  
Stefano Bruno ◽  
Angela Fago ◽  
Roy E. Weber ◽  
Andrea Mozzarelli
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Oxygen Affinity ◽  
Trachemys Scripta ◽  
Oxygen Binding ◽  
Binding Properties ◽  
Cell Variation ◽  
Insight Into

Oxygen-binding properties of single red blood cells from the red-eared turtle Trachemys scripta were measured by microspectrophotometry to describe the variation in oxygen affinity of red blood cells and to gain insight into the distribution of functionally different hemoglobins among red blood cells. Methodologically, this study represents the first report on the cell-to-cell variation in oxygen-binding properties based on oxygen-binding curves of single vertebrate red blood cells. The cells differed significantly with respect to oxygen affinity. Mean oxygen pressure at half saturation of the cells in a blood sample was found to be 20.1 ± 3.3 (SD) Torr. The distribution of oxygen affinities among red blood cells is unimodal, indicating that the two hemoglobins found in turtle blood are not segregated in distinct cells. Therefore, the functional interaction shown by these hemoglobins in vitro is likely to take place in vivo. The considerable variation in oxygen affinity between individual red blood cells calls for its incorporation in models of tissue oxygenation.

Studies of haemoglobin functions by site-directed mutagenesis

1986 ◽  
Vol 317 (1540) ◽  
pp. 443-447 ◽  
Keyword(s):  
Expression Vector ◽  
Oxygen Affinity ◽  
Coagulation Factor ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Oxygen Binding ◽  
Factor X ◽  
Binding Properties ◽  
High Oxygen Affinity ◽  
Alkaline Bohr Effect

Human (3-globin was synthesized in Escherichia coli as a cleavable fusion protein by using the expression vector pLcIIFX|3-globin(nic - ). The authentic (3-globin was liberated by digestion with blood coagulation factor X a and a 2 (3 2 tetramers were reconstituted. The oxygen-binding properties of reconstituted haemoglobin (Hb) were essentially the same as those of human native Hb. Two mutant haemoglobins were constructed by site-directed mutagenesis. HbNymphéas (Cys-93(3->Ser) showed a slightly increased oxygen affinity and diminished co-operativity with normal DPG (2,3-diphosphoglycerate) effect and slightly reduced alkaline Bohr effects. Hb Daphne (Cys-93(3->-Ser, His-143(3->- Arg) showed low co-operativity with high oxygen affinity. The alkaline Bohr effect was slightly reduced, but the DPG effect was enhanced by 50% by the His-143(3^ Arg mutation.

Haemocyanin function in the New Zealand abalones Haliotis iris and H. australis: relationships between oxygen-binding properties, muscle metabolism and habitat

1998 ◽  
Vol 49 (2) ◽  
pp. 143 ◽  
Author(s):  
Rufus M. G. Wells ◽  
John Baldwin ◽  
Scott R. Speed ◽  
Roy E. Weber
Keyword(s):  
Primary Source ◽  
Oxygen Storage ◽  
Oxygen Binding ◽  
Binding Properties ◽  
Muscle Enzyme ◽  
Atp Production ◽  
Muscle Work ◽  
Habitat Differences ◽  
Oxygen Stores ◽  
O2 Binding

Abalone have a well developed capacity for anaerobic metabolism in the pedal musculature, and during exercise the haemolymph oxygen stores may be reserved for more oxygen-dependent tissues. This implies that the haemocyanin-based oxygen transport system is poised towards oxygen storage rather than high rates of oxygen delivery. This proposition was examined by determining O2 -binding properties of haemocyanin from two species of abalone that differ in behaviour and environmental oxygen requirements. Both species showed reversed Bohr and Root effects leading to very high oxygen affinity and reduced cooperativity at low pH. A marked temperature sensitivity of haemocyanin–O2 binding in H. iris (with heat of oxygenation at –55.7 kJ mol-1) impairs oxygen loading above 20˚C and may explain the reduced size and density of populations in warmer northern waters. Muscle enzyme activities indicate low maximum rates of ATP production, with arginine phosphate hydrolysis as the primary source of ATP for elevated levels of muscle work. These data support the hypothesis that oxygen stores support high levels of aerobic muscle work. However, the similarities in enzyme profiles and haemocyanin properties could not be related to behavioural and habitat differences.

pH and haemoglobin oxygen affinity in blood from the Antarctic cod Dissostichus mawsoni

1977 ◽  
Vol 67 (1) ◽  
pp. 77-88
Author(s):  
J. Qvist ◽  
R. E. Weber ◽  
A. L. DeVries ◽  
W. M. Zapol
Keyword(s):  
Molecular Mechanisms ◽  
Oxygen Affinity ◽  
Oxygen Binding ◽  
Low Oxygen ◽  
Binding Properties ◽  
Dissostichus Mawsoni ◽  
Blood Ph ◽  
The Antarctic ◽  
Low Oxygen Affinity

Blood pH in the antarctic cod (Dissostichus mawsoni) and in two Trematomus species, occlrring at --1-9 degrees C, is extremely high (approximately 8-2 to 8-3). This supports and extends Rahn's (1966) model for the temperature-pH relationship in cold-blooded vertebrates. The blood of D. mawsoni shows a low oxygen affinity (P50 approximately equal to 14-5 mmHg at pH 8–16 and −1-9 degrees C). Despite normal in vitro temperature and pH sensitivities, blood P50 increases only slightly when live fish are temperature-stressed (+ 4-0 degrees C), or become acidotic as a result of agitational stress (blood pH 7–71), primarily as a result of compensatory decreases in blood ATP levels. Oxygen-binding properties of ‘stripped’ (cofactor-free) solutions of D. mawsoni haemoglobin were measured in attempts to elucidate the molecular mechanisms involved in the function of the pigment.

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