共查询到20条相似文献,搜索用时 31 毫秒
1.
De Luca G Gugliotta T Scuteri A Romano P Rinaldi C Sidoti A Amato A Romano L 《Cell biochemistry and function》2004,22(3):179-186
The anion influx was measured in order to study the interaction among organic phosphates, magnesium, haemoglobin and the N-terminal of the cytoplasmic domain of band 3 protein in human, chicken and trout erythrocytes. The rate constant for SO(4)(2-) influx in human and trout erythrocytes increased significantly when it was measured with an increased concentration of intracellular Mg(2+). The SO(4)(2-) influx was also measured in human erythrocyte ghosts in the presence and absence of Mg(2+). The smaller activation provoked by Mg(2+) in ghosts could be caused by the presence of a small quantity of haemoglobin which remained inside. The SO(4)(2-) uptake in chicken erythrocytes in the presence and in absence of Mg(2+) was characterized by very similar rate constants. The results suggest that the small increase in intracellular Mg(2+) in the erythrocytes involves an increase in the formation of Mg(2+)-ATP and Mg(2+)-2,3 BPG complexes reducing the affinity of the organic phosphates for Hb. This new situation may influence the functions of the anion transporter with consequent variations of SO(4)(2-) influx throughout the erythrocyte membrane in human and in trout erythrocytes, whereas in chicken RBCs this function cannot occur and, in fact, no increase in sulphate influx was noticeable. The measurement of Hb/O(2) affinity by the use of alternating fixed and variable concentrations of organic phosphates and Mg(2+), confirms the interactions between these elements and their effect on the mechanism of the affinity. When we measured the sulphate influx in the presence of DIDS we found some differences in the three types of cells. 相似文献
2.
Haemoglobin Aalborg (Gly74 (E18)beta----Arg) has a reduced oxygen affinity, in both the absence and the presence of organic phosphates; it has a raised affinity for organic phosphates, and it is moderately unstable. By contrast, haemoglobin Shepherds Bush (Gly74 (E18)beta----Asp) has an increased oxygen affinity in both the absence and the presence of organic phosphates, a diminished affinity for organic phosphates and is also unstable. We have determined the crystal structure of deoxyhaemoglobin Aalborg at 2.8 A resolution and compared it to the structures of deoxy- and oxyhaemoglobin A and of deoxyhaemoglobin Shepherds Bush. The guanidinium group of Arg74(E18)beta protrudes from the haem pocket and donates hydrogen bonds to the E and F helices. The carboxylate group of Asp74(E18)beta forms a hydrogen bond only with residue EF6 and is partially buried, which may be why haemoglobin Shepherds Bush appears to be more unstable than haemoglobin Aalborg. To discover why the latter has a low oxygen affinity, we superimposed the B, G and H helices of haemoglobin A, whose conformation is known to be unaffected by ligand binding, on those of haemoglobin Aalborg. This also brought helices E and the haems into superposition, but revealed a shift of the F helix of deoxyhaemoglobin Aalborg towards the EF-corner. This shift is opposite to that which occurs on ligand binding and on transition to the quaternary oxy-structure, and is linked to an increased tilt of the proximal histidine residue away from the haem axis. Since the relative positions of helices E and F and of the haem group are thought to be the main determinants of the changes in oxygen affinity, the shift of helix F may account for the reduced oxygen affinity of haemoglobin Aalborg. The shift may be due to a combination of steric and electrostatic effects introduced by the arginine residue's side-chain. The effects of the arginine and aspartate substitutions at position E18 beta on the 2,3-diphosphoglycerate affinity are equal and opposite. They can be quantitatively accounted for by the electrostatic attraction or repulsion by the oppositely charged side-chains. 相似文献
3.
Barry J. Frey Roy E. Weber Willie J. van Aardt Angela Fago 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1998,120(4):735-742
The effect of temperature and hypoxic acclimation on the haemoglobin system and intraerythrocytic organic phosphate concentrations in the South African mudfish, Labeo capensis, have been investigated. Exposure to hypoxia or increased temperature raised haemoglobin concentration and decreased NTP/Hb ratio. Temperature acclimation did not effect the oxygenation characteristics of the haemolysate or haemoglobin multiplicity, as evident from isoelectric focussing experiments that showed one cathodic (Hb I) and three anodic haemoglobins (Hb II, III and IV). Oxygen equilibria of the isolated components showed a smaller Bohr effect and lower temperature and organic phosphate sensitivities in the cathodic than in the anodic haemoglobins. Unlike the trout and eel haemoglobin systems, both the anodic and cathodic haemoglobins from L. capensis exhibited sensitivity to organic phosphates but the effect was smaller in the latter. The results indicate that oxygen transport in mudfish blood is supported by variations in the red cell organic phosphate\haemoglobin ratio and the functional differentiation between anodic and cathodic haemoglobins. 相似文献
4.
Studies using equilibrium gel-permeation chromatography demonstrate that formation of the covalent adduct of D-glucose 6-phosphate (G6P) with human haemoglobin promotes dissociation of the haemoglobin tetramer into its component alpha beta dimer pairs [Kdoxy = 2.57 X 10(-6) versus Kdoxy (G6P) = 11.22 X 10(-6) M-haem]. On the other hand, Kd for glucosylated haemoglobin is identical with those of the O2- and CO-liganded forms of intact haemoglobin A0. These data are consistent with the phosphate moiety alone being responsible for a 4.5-fold increase in the tetramer-to-dimer apparent Kd. This suggests the glucose 6-phosphate moiety does not bind to the same sites on haemoglobin as do the free organic phosphates, as suggested by ligand-binding kinetics data or structural analysis. My study presents a working model for studying changes in protein subunit assembly as altered by protein phosphorylations. 相似文献
5.
Embryos from C57BL/6J mice between the gestational ages of 9 and 16 days possess three embryonic haemoglobins EI, EII and EIII, the proportions of which change as a function of gestational age. Component EI, originally present at approx. 65% at day 9, decreases to approx. 20% by day 16, while component EII increases in an inverse manner to that of component EI. During this period component EIII remains essentially constant at approx. 25%. Separation of these species by ion-exchange chromatography has allowed the characterization of the Hill coefficient, Bohr effect, heat of oxygenation and binding of allosterically active organic phosphates for each component. The three components show marked functional heterogeneity and also differ from maternal haemoglobin. Oxygenation curves for whole embryonic blood show distinct deviations from simple binding behaviour. The presence of a high-affinity component within the blood samples may be accounted for by the presence of haemoglobin EI. By using parameters obtained from the study of the isolated components it has been possible to synthesize mathematically the O2-binding curves, obtained experimentally, throughout the gestational period. The characteristics of the isolated haemoglobin components of embryonic mouse blood are discussed in terms of the changing demands for O2 likely to be encountered by the developing embryo. 相似文献
6.
7.
J Oliver T Brittain 《Comparative biochemistry and physiology. B, Comparative biochemistry》1983,76(3):579-583
The haemoglobin purified from a wide range of species undergoes facile sulphaemoglobin formation as well as oxidation. Oxidation by either anions such as azide or organic electron transfer reagents shows similar facility. Facile oxidation of a particular haemoglobin is associated with protection against sulphaemoglobin formation. Oxidation and sulphaemoglobin formation in some instances occur via competing electron transfer reactions. 相似文献
8.
Zn2+ is known to increase the 02 affinity of human haemoglobin. Previous data suggested that Zn2+ exerts its effect by directly binding to haemoglobin, rather than by competing with or binding to 2,3-bisphosphoglycerate. It was also shown that there are two 02-linked zinc-binding sites in haemoglobin, and that Zn2+ does not significantly alter haemoglobin co-operativity or the alkaline Bohr effect. The effect of Zn2+ on 02 affinity of haemoglobin can also be observed for other haemoglobins as diverse as those of cow and chicken. This paper presents new data on the haemoglobin-zinc interaction for normal haemoglobin, des-His146beta-haemoglobin and N-ethylsuccinimide-haemoglobin of humans. For normal haemoglobin (0.05 mM in tetramers), at 20 degrees C in buffer containing 0.1 M-Cl-, 02-dissociation-curve experiments showed that the addition of 0.4-0.5 mM-ZnS04 did not change the Bohr effect between pH 6.71 and 7.29. Similar experiments, with "zinc-ion buffers", showed that the value of the Hill coefficient, h, decreased only slightly if the concentration of free Zn2+ was held constant. For N-ethylsuccinimide-haemoglobin, Zn2+ caused less increase in O2 affinity than for normal haemoglobin. These studies, together with data on the equilibrium binding of Zn2+ to oxy-, deoxy- and des-His146beta-haemoglobins, suggest that zinc is chelated in oxyhaemoglobin by at least three amino acids, two of which are histidine-146beta and cysteine-93beta. 相似文献
9.
The haemoglobin (Hb) of Daphnia magna acclimated to different oxygen conditions was sampled, and in its natively assembled state it was separated by chromatofocusing. The Hb isoforms were analysed for their subunit composition under denaturating conditions by two-dimensional gel electrophoresis. The Hb system is suggested to consist of three predominant Hb aggregates, which are characterised by a specific subunit composition and synthesised in response to different ambient oxygen conditions. In normoxia, a dominant Hb aggregate (DmHbI) with a pI of 4.4-4.6 was composed of subunits B, C, E, F and G. In severe hypoxia, a different dominant Hb isoform (DmHbIII) with a pI of 5.7-5.9 was composed of subunits A, B, C, D, E and F. Further analyses in moderate hypoxia provided evidence for a third Hb isoform (DmHbII) composed of subunits B, C, D, E and F. Sequence alignment and homology modelling of the tertiary structure of the D. magna Hb domains 1 and 2 revealed functionally relevant substitutions of amino acid residues at positions B10, E7 and E11, which determine the functional properties of D. magna haemoglobin in terms of haem contact, oxygen binding and affinity. Both domains are predicted to possess the common haemoglobin fold, but helices C and D are not properly formed, and helix G is interrupted by a short coil. 相似文献
10.
Val AL 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2000,125(4):417-435
Fish are dependent on aerobic metabolism. They respond to changes in oxygen availability by a wide spectrum of compensatory and respiratory adjustments to safeguard tissue oxygenation. Such adjustments are directed to facilitate both oxygen uptake at the gas exchange surfaces and oxygen unloading to tissues. The importance of erythrocytic organic phosphates as regards oxygen transfer has been recognised since 1967 when the 'dramatic' effect of 2,3DPG on human haemoglobin was first reported. The present review examines the appearance of all the major erythrocytic organic phosphates during the evolutionary radiation of fish. In addition, it provides examples illustrating qualitative and quantitative ontogenetic changes of organic phosphates in the red blood cell of several fish species and describes their effects on oxygen affinities. The interaction of the organic phosphates with haemoglobins and divalent cations are also examined. Of particular interest is the regulation of erythrocytic organic phosphates according to both environmental and physiological conditions. 相似文献
11.
《Comparative biochemistry and physiology. A, Comparative physiology》1979,62(1):139-143
- 1. The haemoglobin of the South American lungfishLepidosiren paradoxa has a single component.
- 2. The equilibria of this respiratory protein with oxygen have been investigated both in the blood and with the purified haemoglobin. There is a substantial, normal, alkaline Bohr effect and marked sensitivity to organic phosphates in the haemoglobin solutions.
- 3. Studies on the pH dependence of the kinetics of oxygen dissociation can be interpreted in terms of a normal Bohr effect.
- 4. The kinetics of combination of carbon monoxide have an unusual pH dependence.
- 5. These findings are discussed in terms of the two-state model of Monodet al. (1965)
12.
Differences between horse and human haemoglobins in effects of organic and inorganic anions on oxygen binding.
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B Giardina O Brix M E Clementi R Scatena B Nicoletti R Cicchetti G Argentin S G Condo 《The Biochemical journal》1990,266(3):897-900
Despite the fact that the horse is one of the more common domesticated animals, there are few reports dealing with the properties of its blood, and no comprehensive study has been performed on the reactivity of horse haemoglobin towards organic and inorganic ions. Here we report data on the effects of the organic phosphates D-glycerate-2,3-bisphosphate (2,3-DPG) and InsP6, and of chloride on the properties of horse haemoglobin. Thus the effect of saturating concentrations of 2,3-DPG on the oxygen affinity of horse haemoglobin is about 60% lower than with human adult haemoglobin under the same experimental conditions. The same applies also to InsP6, whose effect on oxygen binding to horse haemoglobin is decreased by about 55% compared with human adult haemoglobin. On the whole, horse haemoglobin appears to be much less sensitive to organic phosphates than previously believed. These results are discussed in the light of the primary structure of the molecule. 相似文献
13.
G Dave 《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》1984,78(2):433-438
The effects of waterborne iron (FeCl3 X 6H2O) on growth, reproduction, survival and haemoglobin content in Daphnia magna were studied from subnormal to toxic concentrations in hard reconstituted water. Low concentrations of iron stimulated reproduction and haemoglobin synthesis after chronic exposure for 21 days. Maximum reproduction occurred between 0.1 and 1 microgram Fe 1(-1). Juvenile growth was not stimulated by iron but was slightly inhibited between 1 and 8 micrograms Fe 1(-1) and above 128 micrograms Fe 1(-1). A slight inhibition of growth persisted for 21 days. Total haemoglobin content was above the control with no waterborne iron at all but one concentration (512 micrograms Fe 1(-1]. The highest value (3.8 X control value) was found at 2 micrograms Fe 1(-1). The haemoglobin content decreased between 64 and 512 micrograms Fe 1(-1) and increased at higher concentrations. The decrease coincided with an inhibited reproduction. The increase was found in non reproductive survivors. A comparison with a previous study in D. magna suggests that ambient conditions (hardness and pH) and ageing of the water are important for the effects of waterborne iron. At a hardness of 250 mg 1(-1) as CaCO3 and a pH range of 7.0-8.0 the ZEP (Zero Equivalent Point) for reproduction was 158 micrograms Fe 1(-1). Continuous exposure to higher concentrations is expected to lead to extinction of a D. magna population. 相似文献
14.
Two generations of red cells (embryonic and definitive), different types of haemoglobins, and special organic phosphates involved in the control of haemoglobin oxygenation (2:3-bisphosphoglycerate, BPG, and inositol-5-phosphate, IP5), have been found progressively during development of the chick. Levels of both organic phosphates, as well as activities of the enzymes involved in BPG synthesis (2:3-bisphosphoglycerate synthase, BPGM) and IP5 formation (phytase), were studied in the erythrocyte populations from embryo, young and adult chickens. Measurement of specific activities of BPGM and phytase in the two subpopulations present in young chickens showed that these phosphates could be specifically and predominantly formed in these two red cell populations. 相似文献
15.
Organotin-protein interactions. Binding of triethyltin bromide to cat haemoglobin. 总被引:1,自引:0,他引:1
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Triethyltin binds to native cat and rat haemoglobin but not to their denatured forms or to other animal haemoglobins. Two molecules of the organotin bind to one molecule of R-state cat haemoglobin with affinity constants of about 1 X 10(5) M-1. Little or no triethyltin is bound to the deoxygenated (T-state) protein. Binding appears to be dependent upon the existence of a specific three-dimensional configuration of cysteine and histidine residues. The properties of the triethyltin-cat haemoglobin complex are consistent with those of a haemoglobin conformer whose allosteric equilibrium is displaced toward the R-state. Its oxygen affinity and rate of oxidation by nitrite is increased, and the rate of reduction of the methaemoglobin derivative by ascorbate is decreased. These effects of triethyltin are opposite and antagonistic to the effects of inositol hexaphosphate. They are exerted on the alpha- as well as beta-haem groups, even though triethyltin is bound at sites on alpha-globin far removed from the haem groups. 相似文献
16.
M Bhattacharyya U Chaudhuri R K Poddar 《International journal of biological macromolecules》1990,12(5):297-301
The interaction of chlorpromazine (CPZ), a widely used antipsychotic tranquillizer, with the allosteric protein haemoglobin, has been studied by different methods. From r versus Cf plot obtained by an equilibrium dialysis experiment, the maximum value of r was found to be 6.8 at 0.15 M NaCl. Binding parameters, namely the affinity constant K and the degree of cooperativity nH, were determined from the Hill plot. Circular dichroism studies indicate a conformation change of haemoglobin in the presence of CPZ. Oxygen has been found to be released from haemoglobin with the progressive addition of CPZ. The extent of the release of oxygen depends on the stoichiometric ratio of CPZ: haemoglobin (D/P). The possible nature of the binding site of the protein has been discussed on the basis of the information obtained from fluorescence measurements. 相似文献
17.
We isolated and sequenced two haemoglobin genes from the early-branching angiosperm Euryale ferox (Nymphaeaceae). The two genes belong to the two known classes of plant haemoglobin. Their existence in Nymphaeaceae supports the theory that class 1 haemoglobin was ancestrally present in all angiosperms, and is evidence for class 2 haemoglobin being widely distributed. These sequences allowed us to unambiguously root the angiosperm haemoglobin phylogeny, and to corroborate the hypothesis that the class 1/class 2 duplication event occurred before the divergence between monocots and eudicots. We addressed the molecular evolution of plant haemoglobin by comparing the synonymous and nonsynonymous substitution rates in various groups of genes. Class 2 haemoglobin genes of legumes (functionally involved in a symbiosis with nitrogen-fixing bacteria) show a higher nonsynonymous substitution rate than class 1 (nonsymbiotic) haemoglobin genes. This suggests that a change in the selective forces applying to plant haemoglobins has occurred during the evolutionary history of this gene family, potentially in relation with the evolution of symbiosis. 相似文献
18.
The dioxygen affinity of Dicrocoelium dendriticum haemoglobin was determined as a function of pH with a thin-layer diffusion technique. From the oxygen dissociation and association curves Hill coefficients h equal 1 were obtained throughout. Ultracentrifugation studies prove this haemoglobin to be monomeric irrespective of pH and ligation state. Thus, Dicrocoelium haemoglobin is a non-cooperative monomer. It has the highest O2 affinity so far known for any monomeric haemoglobin: its half-saturation pressure, p50 value, ranges at 25 degrees C from 0.016 mm Hg to 0.15 mm Hg (2.13-20.0 Pa) dependent on pH. Dicrocoelium haemoglobin shows an acid Bohr effect only and as such it constitutes a new class of haemoglobins. Its log p50 versus pH plot (Bohr effect curve) is characterized by a large amplitude, delta log p50 = 0.96, and an inflection point (Bohr effect pK) at pH 5.0. A model for the acid Bohr effect of D. dendriticum haemoglobin is proposed. By generalization, both the alkaline and the acid Bohr effect in various monomeric haemoglobins may arise from a single Bohr group complex (salt bridge). 相似文献
19.
We have studied the effects of organic cosolvents (monohydric alcohols and formamide) on the oxygen equilibrium of bovine haemoglobin and have compared them with the effects of the same cosolvents on the oxygen equilibrium of human haemoglobin. Our results indicate: (1) that in agreement with previous suggestions, the lower affinity of bovine haemoglobin for oxygen is not due to an increased number of salt bridges stabilizing the T structure; (2) that, following T----R transition, more hydrophobic surface is exposed to the solvent by bovine than by human haemoglobin. We suggest, therefore, that a relevant role in keeping the oxygen affinity of bovine haemoglobin lower than that of human haemoglobin is played by the higher free energy needed to expose this more hydrophobic surface to the solvent. We stress, however, that our analysis does not enable us to say which particular amino acid residues are concerned in these effects. 相似文献
20.
Des arginine 141 a haemoglobin (the haemoglobin in which the C-terminal arginine of the a chain has been removed) has a high affinity for oxygen and a reduced co-operativity in its oxygen equilibrium binding. The kinetic consequences of this modification are investigated in this paper. Deoxy des Arg haemoglobin binds carbon monoxide faster than does haemoglobin A, whilst oxy des Arg haemoglobin loses oxygen more slowly. These results are correlated with the oxygen equilibrium binding properties of des Arg haemoglobin. The carbon monoxide binding kinetics have been interpreted as implying a change in the parameter c (of the allosteric model), as well as L, when this arginine is removed from haemoglobin. 相似文献