共查询到20条相似文献,搜索用时 34 毫秒
1.
The kinetics of human oxyhemoglobin coagulation in neutral phosphate buffer in the presence of mercury acetate at 20 degrees has been studied using turbidimetric methods. The addition of small amounts of concentrated Hg2+ solution leads to rapid local protein coagulation with subsequent dissolution of the formed coagulate. Coagulation can be inhibited by addition of Tris that binds to mercury ions. The pattern of oxyhemoglobin coagulation is determined by molar Hg2+/protein ration rather than by total Hg2+ concentration. 相似文献
2.
The initial velocity of coagulation of human oxyhemoglobin in tris-HCl buffer measured by turbidimetric method, pH 7.2 in the presence of phenylmercuryacetate made it possible to estimate the amount of moles of this reagent stechiometrically binding with hemoglobin without coagulation of the latter. At 15-30 degrees C this amount is 30-34 mole per hemoglobin-tetramer. At temperature increase from 30 to 42.5 degrees C the amount of the reagent necessary for protein coagulation sharply decreases. A model is proposed assuming that oxyhemoglobin coagulation proceeds only during binding of the reagent with specific protein sites. 相似文献
3.
A E Myshkin 《Izvestiia Akademii nauk SSSR. Seriia biologicheskaia》1992,(1):90-102 concl
The results are discussed of studies on oxyhemoglobin coagulation in neutral phosphate buffer and acidic acetate buffer at pH ranging from 5.85 to 4.90. Peculiarities are shown of the effect of strong complexon on the oxyhemoglobin-coagulum-mercuric acetate system in neutral tris-buffer. Coagulation characteristics are cited for polymeric oxyhemoglobin in presence of mercury ions. 相似文献
4.
By the dynamics of human oxyhemoglobin coagulation in the presence of phenyl mercury acetate in tris-AcOH buffer, pH 7.2 the number of moles of PhHg+ stechiometrically bound with protein at different temperatures was estimated. Within the temperature range 15-30 degrees C this value is constant--32-34 mole per 1 mole of HBO2-tetramer. Within the range 30-40 degrees C it rises to approximately 40. Coagulation of oxyhemoglobin modified with PhHg+ cation is reversible in contrast to HBO2 coagulation modified with uncharged PhHgCl. 相似文献
5.
Coagulation of bovine oxyhemoglobin in the presence of mercuric acetate in concentrations within a range including concentrations exceeding those required to block the single pair of thiol groups of the protein has been investigated in Tris-acetate buffer. The values of initial coagulation rate plotted against mercury-to-hemoglobin molar ratio give curves exhibiting a clear break points at ratios corresponding to full blocking of the mentioned thiol groups. Larger amounts of mercury reagents producing enhanced protein coagulation effect depend approximately quadratically on the mercury concentration. Interaction of the excess mercuric ions with some mercury-binding sites located on or near the dimer-dimer contact surfaces of the protein producing stronger coagulation effect is suggested. 相似文献
6.
Histidin has been shown to effectively inhibit coagulation of horse oxyhemoglobin (HbO(2)) modified by mercury(II) ion bound to reactive thiol groups of protein. Kinetic parameters were measured and the histidin-to-mercury binding constant was kinetically estimated. Histidin, as other pharmaceutically acceptable compounds with some mercury-binding capacity, has been suggested to alleviate mercury intoxication conditions. 相似文献
7.
A E Myshkin 《Izvestiia Akademii nauk SSSR. Seriia biologicheskaia》1991,(4):583-593
An analytical review of studies on human oxyhemoglobin coagulation has been performed by the author jointly with V. S. Koniaeva and L. D. Bogdanova within a period from 1985 to 1990. It was shown that the oxyhemoglobin coagulation modified by mercurials proceeded without any essential alteration of native protein conformation. A hypothesis is discussed that the oxyhemoglobin coagulation results from the primary polyaggregation of dimer fragments and that hydrophobic sites which provide for dimer-to-dimer contacts in native tetrameric oxyhemoglobin, participate in this process. 相似文献
8.
The number of Bohr protons released upon oxygenation has been measured over a large range of human hemoglobin concentrations (0.02 to 4.5 mM) in the presence of equimolar amounts of D-glycerate 2,3-bisphosphate. From these data the association constants for the binding of this organic phosphate to deoxyhemoglobin and oxyhemoglobin were calculated at different pH values. The maximum number of protons absorbed upon binding to oxyhemoglobin was determined as well. The maximum number of protons bound to deoxyhemoglobin upon binding of D-glycerate 2,3-bisphosphate was measured independently. From the pH dependence of the association constants and the maximum number of protons absorbed it could be concluded that only one D-glycerate 2,3-bisphosphate can be bound to both deoxyhemoglobin and oxyhemoglobin. 相似文献
9.
pH-dependent absorption in the B and Q bands of oxyhemoglobin and chemically modified oxyhemoglobin (BME) at low Cl- concentrations.
下载免费PDF全文
![点击此处可从《Biophysical journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
We have measured the optical absorbance in the maxima of the Q and B bands for oxyhemoglobin and oxyhemoglobin (BME) in dependence on the pH value of the solution in the region between pH 4.4 and pH 10. From the absorbance data optical titration curves are derived for both bands. These yield for oxyhemoglobin pK values 4.3, 5.3, 6.8, 7.8, and 9.0, whereas for oxyhemoglobin (BME) only one pK value at 4.3 is observed. These data are in good agreement to those derived recently from resonance Raman spectroscopy. The changes of the oscillator strengths in the Q bands are interpreted in terms of Gouterman's four-orbital model to arise from A1g-distortions of the heme group, resulting from changes of the heme-apoprotein interactions due to protonation processes of amino acid-side groups in the beta-chains. The difference between the sets of pK values in oxyhemoglobin and oxyhemoglobin BME is explained from the fact that the bifunctional reagent BME blocks important pathways of heme-apoprotein interactions. The fact that in any case increase of the Q band absorbance is accompanied by a corresponding increase in the B band absorbance leads us to the conclusion that the electronic structure of the B bands has to be described in terms of a six-orbital model, taking into account configurational interaction with the L and N bands. 相似文献
10.
Thrombelastography (TEG) is a method that is used to conduct global assays that monitor fibrin formation and fibrinolysis and platelet aggregation in whole blood. The purpose of this study was to use a well-characterized tissue factor (Tf) reagent and contact pathway inhibitor (corn trypsin inhibitor, CTI) to develop a reproducible thrombelastography assay. In this study, blood was collected from 5 male subjects (three times). Clot formation was initiated in whole blood with 5 pM Tf in the presence of CTI, and fibrinolysis was induced by adding tissue plasminogen activator (tPA). Changes in viscoelasticity were then monitored by TEG. In quality control assays, our Tf reagent, when used at 5 pM, induced coagulation in whole blood in 3.93 ± 0.23 min and in plasma in 5.12 ± 0.23 min (n=3). In TEG assays, tPA significantly decreased clot strength (maximum amplitude, MA) in all individuals but had no effect on clot time (R time). The intraassay variability (CVa<10%) for R time, angle, and MA suggests that these parameters reliably describe the dynamics of fibrin formation and degradation in whole blood. Our Tf reagent reproducibly induces coagulation, making it an ideal tool to quantify the processes that contribute to mechanical clot strength in whole blood. 相似文献
11.
A significant difference was discovered in low temperature (77 K) gamma-radiolytic behavior of 20% aqueous solutions of human oxyhemoglobin and partly denaturated methemoglobin. In the latter case twice as high yield of the sum of free radicals and OH radicals was observed, as well as presence in the ESR spectrum of a narrow singlet line at g 2.00 (absent for irradiated solutions of oxyhemoglobin) ascribed to the stabilized electron. 相似文献
12.
The efficiency of in vivo blood irradiation by a laser light source (λ = 632.8 and 670 nm) and a mercury lamp (UV light, λ = 254 nm) was compared. Absorption spectra, gas content, oxyhemoglobin content, hemoglobin oxygen saturation, concentrations of lactate and glucose were studied for both irradiated and control samples. Hemoglobin was assumed to be the primary photoacceptor of light radiation for the indicated wavelengths. No substantial differences have been found between the effects of laser and non-laser irradiation. We conclude that the biological impact of the procedure is related to photoinduced changes in hemoglobin oxygen saturation. 相似文献
13.
Interaction of acetylphenylhydrazine with oxyhemoglobin A in a hemolysate or in intact red cells resulted in the formation of ferrihemochromes as shown by a characteristic optical spectrum. The same optical spectrum was observed in a suspension of red cell ghosts containing numerous Heinz bodies. Electron paramagnetic resonance of actylphenylhydrazine-incubated red cells disclosed the presence of previously identified reversible ferrihemochromes, which can be reduced to functional hemoglobin, and irreversible ferrihemochromes, which cannot be reduced to functional hemoglobin. (Ferrihemochromes are defined as low spin forms of ferric hemoglobin having heme ligands endogenous to the protein structure). In contrast, only irreversible ferrihemochromes could be observed in ghosts containing Heinz bodies. In addition both optical and magnetic features of sulfhemoglobin were observed in an acetylphenylhydrazine-treated red cell hemolysate. Similar optical features are produced by the interaction of aromatic nitrogen-containg reductants with purified oxyhemoglobin in the presence of (NH4)2S. This reaction is not effected by the presence of catalase, suggesting that H2O2 is not an intermediate of the reaction. It is concluded that the mechanism of action of acetylphenylhydrazine with oxyhemoglobin is two-fold, ultimate reduction to high spin ferric hemoglobin followed by ferrihemochrome formation. Thus it appears that the pathway of denaturation of hemolytic anemias and thalassemia or induced by chemical reagents, entails a common route involving the formation of ferric hemoglobin by a reductive mechanism, followed by reversible ferrihemochromes, irreversible ferrihemochromes, and ultimately, precipitation. 相似文献
14.
Human oxyhemoglobin is converted to methemoglobin by a wide array of organic and inorganic reductants. Depending upon the concentration and nature of the reductant, varying amounts of deoxyhemoglobin are produced. The general overall sequence is: FeO2 leads to (1) FeIII leads to (2) FeII. The intermediacy of methemoglobin can be demonstrated by direct spectral observation and by cyanide trapping. For organic reductants, the second-order rate constants for (1) vary from greater than 300 (phenylhydroxylamine) to 1.4 X 10(-4) M-1 s-1 (malononitrile). Generally the rates parallel the ease of hydrogen abstraction by iron-bound oxygen from the substrate, and simply hydrocarbons are reactive. Rates for these processes have been ascertained with recrystallized protein, lysed cells, and intact human erythrocytes. At room temperature oxyhemoglobin quantitatively converts benzaldehyde to benzoic acid and hydroquinone to benzoquinone. Rates for inorganic species (process 1) range from greater than 7 X 10(3) (chromous ion) to 0.015 M-1 s-1 (ferrocyanide). Ferrous ion rapidly deoxygenates oxyhemoglobin by direct attack on the oxy complex but methemoglobin is not an intermediate with this reagent. Taken together the results support the theoretical prediction that reductants should oxidize oxyhemoglobin, and they demonstrate at least some degree of radical character to the oxy complex. 相似文献
15.
L Forlani C Ioppolo L Civalleri D L Currell M A Marini 《Journal of inorganic biochemistry》1986,26(4):289-294
Calorimetric studies of the effect of superoxide dismutase and/or catalase on the reduction of dioxygen into water by dithionite in oxyhemoglobin have been carried out and the results compared with those in red cell hemolysates. In the absence of the enzymes the stoichiometry (moles dithionite/mole dioxygen) is less than the value of 2:1 which was found previously in red cell hemolysates [Forlani et al., J. Inorg. Biochem. 20, 147-155 (1984)]. In the presence of either superoxide dismutase or catalase alone the stoichiometry increases but is still less than 2:1. In the presence of both enzymes the stoichiometry and the shape of the thermogram is that previously observed for hemolysates, suggesting the presence of a hemoglobin-catalase-superoxide dismutase integrated system. The absence of a calorimetric signal for hydrogen peroxide in the reduction of oxyhemoglobin in the presence of superoxide dismutase suggests a wider biological role of superoxide dismutase than previously thought. 相似文献
16.
V Iu Titov Iu M Petrenko V A Petrov Iu A Vladimirov 《Biulleten' eksperimental'no? biologii i meditsiny》1991,112(7):46-49
The process of oxyhemoglobin oxidation initiated by hydrogen peroxide in low (10(-7) M) concentrations was investigated. It was found, that H2O2 in this concentration is able to induce the process of chain oxidation of oxyhemoglobin to methemoglobin. The following observations indicate that the process is essentially the chain reaction: 1) The amount of the methemoglobin in haem groups, produced in the reaction, exceed by 20 times the quantity of hydrogen, added initially, to induce the oxidation. 2) Catalase stopped this process at any stage of the reaction. This fact implies that the chain process involves generation of new molecules of H2O2 in the course of oxidation of oxyhemoglobin. The chain reaction proceeded only in the presence of oxygen. But if oxygen was introduced into hemoglobin solution, preincubated with H2O2 in vacuum, than again the oxidation of hemoglobin developed. Apparently, H2O2 in low concentrations appears, mainly, as an inductor of the oxyhemoglobin autooxidation. 相似文献
17.
The elastic modulus (G') of factor XIIIa induced fibrinogen gels was found to be substantially lower than the G' of fibrin gels that were formed by clotting fibrinogen with thrombin. The addition of fibronectin and/or the reducing reagent dithiothreitol (DTT) to the factor XIIIa coagulation mixture led to the formation of a weaker gel structure, while the rigidity of thrombin induced clots was not appreciably affected by the inclusion of the DTT but increased somewhat in the presence of fibronectin. The reasons for the differing clot rigidities are discussed in terms of biochemical mechanisms. 相似文献
18.
Factor VIII-light chain (FVIII-LC) and Factor VIII-heavy chain (FVIII-HC) were isolated separately from human plasma and were without coagulation activity. When FVIII-LC and FVIII-HC preparations were mixed, coagulation activity was generated in the presence of Mn2+, Ca2+, or Co2+. Mn2+ was most effective and with Ca2+ maximal activity was first obtained after 8 days. Bovine Factor X (FX) accelerated recombination and was able to increase the amount of FVIII:C generated up to 10-fold when FVIII-LC/HC were incubated with Ca2+ for 20 h. When recombination was performed in the presence of micromolar concentrations of sulfhydryl reagents, the total amount of FVIII:C generated was increased up to 4-fold and in excess of FVIII-HC it was possible to drive 70% of FVIII-LC into active complex. If FVIII-HC was prepared in the presence of a sulfhydryl reagent it was possible to drive 75% of FVIII-HC into active complex with FVIII-LC in excess. Me2+, which is necessary for recombination, catalyzes disulfide formation, and it is proposed that FVIII subunits have free sulfhydryl groups. The presence of sulfhydryl reagents during purification of FVIII-HC and during recombination retains the individual subunits in a conformation suitable for recombination. 相似文献
19.
Lisauskas SF Cunha NB Vianna GR Mendes EA Ramos GL Maranhão AQ Brígido MM Almeida JO Baptista HA Motta FL Pesquero JB Aragão FJ Rech EL 《Biotechnology letters》2008,30(12):2063-2069
Human factor IX is synthesized in the liver and secreted in the blood, where it participates in a group of reactions involving
coagulation factors and proteins that permit sanguinary coagulation. In this work two lines of transgenic mice were developed
to express the FIX gene in the mammalian glands under control of milk β-casein promoter. The founding females secreted the FIX in their milk
(3% total soluble protein). The stable integration of transgene was confirmed by southern blot analysis. The presence of the
FIX recombinant protein in the milk of transgenic females was confirmed by western blot and the clotting activity was revealed
in blood-clotting assays. The coagulation activity in human blood treated with recombinant FIX increased while the time of
coagulation decreased. Our results confirm the production of a large amount of recombinant biologically active FIX in the
mammary gland of transgenic mice. 相似文献
20.
Studies on the mechanism of the stimulation of polymerase II-catalyzed RNA synthesis by mercury compounds 总被引:1,自引:0,他引:1
The specific stimulation of alpha-amanitin-sensitive RNA synthesis in isolated nuclei by methyl mercury (Frenkel, G. D., and Randles, K. (1982) J. Biol. Chem. 257, 6275-6279) has been further investigated. Using the method of alkaline hydrolysis/uridine analysis to determine the number of RNA chains growing in vitro, it was found that the stimulation could not be accounted for by an increase in the number of growing chains. The stimulatory effect of heparin (Coupar, B. E. H., and Chesterton, C. J. (1977) Eur. J. Biochem. 79, 525-533), was found to be additive with that of methyl mercury at saturating concentrations of the latter. Various detergents were found to affect RNA synthesis per se and to modify the stimulatory effect of methyl mercury, suggesting that the stimulation by methyl mercury requires a degree of structural integrity of some nuclear components. The ability of a number of other mercury compounds to stimulate RNA synthesis was investigated. None of the inorganic compounds examined, i.e. HgCl2, HgSO4, and Hg(ClO4)2, stimulated synthesis. Among the alkyl organic compounds tested in addition to methyl mercury, ethyl mercury also stimulated RNA synthesis, but dimethylmercury did not. Among the aryl compounds tested, phenylmercury did not stimulate synthesis whereas p-hydroxymercuribenzoate and p-hydroxymercuribenzenesulfonate did. N-Ethylmaleimide, a nonmercurous sulfhydryl reagent, was found to have only weak ability to stimulate RNA synthesis, compared to a comparable mercury-containing sulfhydryl reagent, p-hydroxymercuribenzoate. The stimulatory effect of the latter was, however, effectively competed out by the former, indicating that sulfhydryl binding is necessary for the stimulation but not sufficient. This conclusion was reinforced by experiments which utilized a model system to measure the ability of various mercury compounds to compete with N-ethylmaleimide in binding to cysteine. The results showed that even compounds such as phenylmercury and the inorganic mercurials, which are unable to stimulate RNA synthesis, are able to bind to a sulfhydryl group. 相似文献