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A Congiu-Castellano A Bianconi M Dell'Ariccia S Della Longa A Giovannelli E Burattini M Castagnola 《Biochemical and biophysical research communications》1987,147(1):31-38
The X-ray absorption near edge structure (XANES) spectra of the human adult and foetal hemoglobin, of the isolated alpha and beta chains, in the oxygenated forms, and of the oxymyoglobin and carp oxyhemoglobin have been measured at the wiggler beam line of the Frascati Synchrotron radiation facility. The bonding angle of oxygen molecule at the iron site in these hemoproteins in solution, has been measured using the multiple scattering theory for data analysis. 相似文献
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Samples of 90% enriched 57Fe hemoglobin and its isolated subunits have been prepared. M?ssbauer spectroscopic measurements have been made on three such samples. Sample one contained contributions of oxyhemoglobin, deoxyhemoglobin, and carbonmonoxyhemoglobin. This sample was studied from a temperature of 90 K down to 230 mK. Measurements were also made at 4.2 K using a small applied magnetic field of 1.0 T. In general, the measured quadrupole splittings and isomer shifts for each component agreed with previous measurements on single component samples in the literature, and thus demonstrated that chemically enriched hemoglobin has not been altered. The second and third samples were isolated alpha and beta subunits, respectively. We have found measurable M?ssbauer spectral differences between the HbO2 sites in the alpha subunit sample and the beta subunit sample. The measured M?ssbauer spectral areas indicate that the iron ion has the largest mean-square displacement at the deoxy Hb sites as compared to that at the oxy- and carbonmonoxy Hb sites. The mean-square displacement at the HbO2 sites is the smallest. 相似文献
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The similarities in the Mossbauer spectra of Hb and its α and β subunits, both in the oxygenated and deoxygenated states, were observed. The relative intensities of the absorption dips in the Mossbauer spectra of 57Fe in anhydrohemoglobin (AHb) and anhydrous separated alpha (Aα) and beta (Aβ) chain samples were found to be very sensitive to the conditions of samples preparation. This indicates the formation of hemochromogen as an impurity in the absorbers during the process of preparation. When contributions due to the impurity in the samples were subtracted, Mossbauer spectra of 57Fe in AHb, Aα, and Aβ remained similar. Therefore, the electronic structure of the iron cation is the same for Hb and its subunits in the anhydrous state also. 相似文献
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B Campbell C Fronticelli A Zachary E Bucci 《The Journal of biological chemistry》1986,261(9):3931-3933
Apohemoglobin S formed a gel in the cold (5 degrees C) with a protein concentration in the supernatants after centrifugation of the gels (Csat) near 27 g/dl, in 0.02 M phosphate buffer at pH 7.2. Under the same experimental conditions in mixtures of apohemoglobin S and deoxyhemoglobin S the solubility of hemoglobin S in the cold was decreased from Csat greater than 40 g/dl in the absence to about 18 g/dl in the presence of apohemoglobin S. Conversely, in the same mixture, Csat of apohemoglobin S was decreased to about 5 g/dl. Also, gelling occurred in mixtures of oxyhemoglobin S and its apoderivative. Apohemoglobin A alone did not form gels; however, it induced fiber formation in deoxyhemoglobin S in the cold; unlike apohemoglobin S, it was not included in the precipitate. Gels of apohemoglobin S were not birefringent, and inspection at the electron microscope failed to show the presence of organized structures. Excluded volume effects were probably at the origin of the decreased solubility of hemoglobin S and apohemoglobin S in the presence of each other. 相似文献
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Summary. Although peroxynitrite is believed to be one of the most efficient tyrosine-nitrating species of biological relevance so far identified, its nitration efficiency is nevertheless limited. In fact, the nitrating species formed through peroxynitrite decay are caged radicals (OH/NO2 or, in the presence of carbon dioxide, CO3
–/NO2) and the fraction that escapes from the solvent cage does not exceed 30–35%. One exception may be represented by metal-containing compounds that can enhance the formation of nitrotyrosine through a bimolecular reaction with peroxynitrite. Moreover, if the metal is also regenerated in the reaction, the compound is considered a nitration catalysts and the yield of tyrosine nitration enhanced several fold. Examples of peroxynitrite-dependent nitration catalysts are the Mn-superoxide dismutase, some cytochromes and several metalloporphyrins. On the contrary, it has been claimed that some hemoproteins are scavengers of peroxynitrite and play a role in limiting its biodamaging and bioregulatory activity. In this review, we discuss the case of hemoglobin, which is probably the major target of peroxynitrite in blood. This protein has been reported to protect intracellular and extracellular targets from peroxynitrite-mediated tyrosine nitration. This property is shared with myoglobin and cytochrome c. The possible mechanisms conferring to these proteins a peroxynitrite scavenging role are discussed.Present address: Laboratorio di Tossicologia Applicata ed Ecotossicologia, Istituto Superiore di Sanità, Rome, Italy. 相似文献
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Optically active absorption bands of hemoglobin and its subunits 总被引:2,自引:0,他引:2
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Heterogeneity of the isolated subunits of the fetal and adult human hemoglobin in solution, detected by XANES spectroscopy 总被引:1,自引:0,他引:1
A Congiu Castellano M Castagnola E Burattini M Dell'Ariccia S Della Longa A Giovannelli P J Durham A Bianconi 《Biochimica et biophysica acta》1989,996(3):240-246
Differences in the local structure of the heme in the isolated alpha-, beta- and gamma-chains of the adult and fetal human hemoglobin are detected by XANES (X-ray absorption near-edge structure) spectroscopy. The ligand bonding angle to the iron ion in the ligated forms and the displacement of the Fe respect to the porphyrin plane in the deoxy forms are found to be different for each chain. 相似文献
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The geminate recombination of CO with carboxy hemoglobin (Hb4(CO)3) following a ten nanosecond laser pulse and the overall combination of the fourth CO with Hb4(CO)3 has been studied as a function of pH in the presence and absence of inositol hexaphosphate. The results indicate that the kinetics of both reactions are independent of pH and phosphate concentration. The results are discussed in terms of a two-step mechanism: a pre-equilibrium step followed by heme—ligand bond formation. The latter is also known as the geminate recombination reaction (Hb + CO α Hb · CO α HbCO). 相似文献
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Studies on the status of tyrosyl residues in cobrotoxin 总被引:3,自引:0,他引:3
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Gene A. Homandberg 《Biochemical and biophysical research communications》1982,105(3):1109-1114
Maleic anhydride modification of tetrameric mouse β-glucuronidase, followed by a 70° incubation, dissociated the tetramer into inactive monomers. Deblocking of this derivative allowed 80% regeneration of activity and tetrameric structure. The enzyme was inactivated by reaction with N-ethyl-S-phenylisoxazolium-3′-sulfonate, tetranitromethane and succinic anhydride, but not when a competitive inhibitor was added to enzyme prior to modification. These data suggest that the active site residues of mouse β-glucuronidase include carboxyl, tyrosyl and lysyl residues. In comparison, it has been reported that rat β-glucuronidase is inactivated by chemical modification of carboxyl, tyrosyl and residues. 相似文献
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Fluorescent conjugates of hemoglobin A, its isolated β-chain, and the apo-derivative of the β-chain have been prepared in which the β-93 sulfhydryl was conjugated with 1,5-AEDANS. Radiationless enery transfer to the heme group results in a major decrease in fluorescence intensity and decay time. Measurements of the time decay of fluorescence anisotropy, employing single-photon counting, indicate that the apparent rotational correlation time is, in each case, substantially reduced from the value expected for a rigid molecule of the same molecular weight. This observation raises the possibility that internal degrees of rotational freedom exist. 相似文献
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G Boguta A M Dancewicz 《International journal of radiation biology and related studies in physics, chemistry, and medicine》1983,43(3):249-265
Insulin, ribonuclease, papain and collagen solutions saturated with nitrogen, N2O or air were irradiated with doses of 10 to 640 Gy of gamma rays. Protein solutions were also oxidized enzymatically in a system of horse-radish peroxidase: hydrogen peroxide. Column chromatography (Sephadex G-75 or Sephacryl S-200) of treated protein solutions revealed that they contain protein molecular aggregates. Nitrogen saturation of solution before irradiation was most favourable for radiation-induced aggregation of proteins. Fluorescence analysis of protein solutions resulted in detection of dityrosyl structures in irradiated as well as in enzymatically oxidized proteins. Concentration of dityrosine in proteins studied was determined fluorimetrically in their hydrolysates separated on BioGel P-2 column. In irradiated proteins, dityrosine was present almost exclusively in their aggregated forms. In proteins oxidized enzymatically, dityrosine was also present in fractions containing apparently unchanged protein. Mechanisms which could account for differences in the yield of dityrosine formation in radiolysis and in enzymatic oxidation of proteins are suggested. 相似文献