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A Takeda

Publications and source records attributed to A Takeda.

At least 541 records · Page 30Linked to original sources
PubMed1981

On the acid denaturation of porcine erythrocyte catalase in relation to its subunit structure.

Porcine erythrocyte catalase [EC 1.11.1.6] (molecular weight, ca. 250,000) was found to dissociate partially between pH 3.5 and 3.0 and completely below pH 3.0 into two presumably identical 1/2-sized subunits (molecular weight, 112,000) as estimated by ultracentrifugal analyses. This dissociation was accompanied by a marked change in hydrodynamic properties; the sedimentation coefficient decreased from about 11S to 4S. This acid denaturation also resulted in complete loss of enzyme activity and disappearance of absorption bands characteristic of heme protein, in particular, a shift of the Soret band from 405 nm to a small and broad band at 375 nm. The change in enzyme activity correlated well with that of the Soret band, depending on the denaturation time and pH used. Reversible recovery of enzyme activity was not detected below pH 3.1 after 2 h denaturation. The pH dependence of alpha-helical content estimated from the CD intensity at 222 nm also correlated well with that of enzyme activity. The rate constants of initial reaction of acid denaturation at several pHs were determined by following the changes in the Soret band with time, since the changes showed an isosbestic point at 384 nm. The results revealed that the first-order rate constant at pH 3.0 was 45 times larger than that at pH 3.4, indicating that the rate of acid denaturation increased rapidly within a narrow acidic pH range. The temperature dependence of the denaturation rate was also measured and the activation energy for the acid denaturation was found to be 76.3 kcal/mol from an Arrhenius plot.

Animals↗
PubMed1981

[Labeling of unnatural amino acids with 99mTc and tissue distribution of the labeled products in mice].

The selective affinity of amino acids for pancreas, due to its high rate of protein synthesis, has prompted us to synthesize new radio-labeled amino acids for pancreatic imaging. Labeling of 19 unnatural amino acids with 99mTc was tried in the usual way, and 10 labeled products were obtained with low yields. The highest distribution of radioactivity from all these labeled amino acids, which had been injected intravenously into mice, was found in the kidney, whereas little activity was found in the pancreas. Thier low distributions in stomach suggest little dissociation of the labeled amino acids into 99mTcO4- in mice. It is presumed that these labeled amino acids behave unlike their original acids in the body.

Amino Acids↗
PubMed1980

The interactions of thiol compounds with porcine erythrocyte catalase.

The effects of thiol compounds on the conformation of porcine erythrocyte catalase were examined. The thiol compounds showed two types of reactivity with the catalase in terms of changes in absorption spectra. One is characterized by the appearance of a new absorption maximum at 595 nm; this was seen with 2-mercaptoethanol (designated as inactive catalase type I). The other is characterized by new maxima at 535 and 570 nm, and this was seen with reduced glutathione, dithiothreitol, cysteine, and cysteamine (inactive catalase type II). The thiol compounds caused gradual inactivation of catalase, correlating with the enhancement of the absorption maximum at 595 nm or 570 nm. Removal of excess thiol reagents from the reaction mixtures caused partial recovery of activity, which was more marked with inactive catalase type II. Similar reversibility was observed in the absorption, CD and MCD spectra, whereas reversibility was not observed for inactive catalase type I. The MCD spectra suggested conversion of heme groups from a high to a low spin state on incubation with thiols, e.g., reduced glutathione, leading to inactive catalase type II. alpha-Helical conformation of the polypeptide backbone and titratable free SH groups in the catalase molecule were unaffected by all these thiol treatments. It is suggested that "active oxygen" which may be produced on incubation of catalase with thiol compounds, was responsible for the formation of inactive catalase type II.

Animals↗
PubMed1978

Studies on the negative circular dichroic bands around 297 nm of ribosomes from bacterial cells.

The small negative CD bands around 297 nm of isolated 30-S and 50-S ribosomal subunits were precisely measured for three bacteria, Bacillus stearothermophilus, Bacillus subtilis and Escherichia coli Q 13. The intensities of the negative CD bands of 30-S subunits were always much greater than those of 50-S subunits irrespective of the bacterial strains, which may be related to the difference in comformations of rRNAs and proteins in the complexes between these subribosomal particles. The dissociation of 70-S ribosomes into two subunits by lowering Mg2+ concentration caused evident enhancement of intensity of the 297 nm CD band, which was completely reversed by the association of the two subunits into 70-S particles. The melting profiles of CD spectra 3 B. stearothermophilus and E. coli were compared and both subunits of the former were found to be more heat stable than those of the latter. It was found that 5 M urea and 0.5% sodium dodecyl sulfate (SDS) treatment caused considerable reduction of the negative CD intensity around 297 nm of 30-S subunits but no significant change of 50-S subunits, while no significant change was observed for the CD spectra of isolated 16-S and 23-S rRNAs by the same treatment. Effects of EDTA treatment and then addition of Mg2+ on the CD spectra and fluorescence emission spectra of the subunits were also observed and the contribution by the interaction between rRNA s and proteins in ribosomes to the small negative band around 297 nm was discussed.

Bacillus subtilis↗
PubMed1977

On the specific association of porcine erythrocyte catalase caused by formation of disulfide cross-links.

Porcine erythrocyte catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase, EC 1.11.1.6) has been purified from porcine blood by DEAE-cellulose column chromatography, ammonium sulfate fractionation and CM-cellulose column chromatography. The purified enzyme was found to associate into larger molecules than the native one when it was stored at 4degrees C for more than one week. The associated molecules can be detected by gel filtration on a Bio-gel A-1.5 m column and disc gel electrophoresis as well as ultracentrifugal analysis. Molecular weights of the associated catalase molecules were about 500 000, 750 000, 1 000 000 and so forth estimated by gel filtration and disc gel electrophoresis, corresponding to dimer, trimer and tetramer respectively, of a native molecule (monomer) with a molecular weight of about 250 000. The association of catalase molecules is found to be time-dependent and to proceed seemingly from monomer through dimer as an intermediate. From the effects of several thiol reagents or reducing reagents on the association process and spectrophotometric titration of SH groups, it is inferred that this specific association of procine erythrocyte catalase is caused by formation of intermolecular disulfide cross-links due to air oxidation of SH groups in the protein moiety.

Animals↗
PubMed1977

Model for "charge-relay": acceleration by carboxylate anion in intramolecular general base-catalyzed ester hydrolysis by the imidazolyl group.

The effect of benzoate anion on intramolecular general base-catalyzed ester hydrolysis by the imidazolyl group in endo-5-[4'(5')-imidazolyl]bicyclo[2.2.1]hept-endo-2-yl trans-cinnamate was examined in dioxane/H(2)O solutions. Benzoate anion exhibited a remarkable acceleration of the intramolecular general base-catalyzed hydrolysis of endo-5-[4'(5')-imidazolyl]bicyclo[2.2.1]hept-endo-2-yl trans-cinnamate by the imidazolyl group. The rate of hydrolysis in the presence of the benzoate anion increased with the dioxane mole fraction and was proportional to the concentration of benzoate anion. On the other hand, the rate of hydrolysis of endo-5-[4'(5')-imidazolyl]bicyclo[2.2.1]hept-endo-2-yl trans-cinnamate in the absence of benzoate anion decreased with the dioxane mole fraction. Thus, the ratio of the rate in the presence of benzoate anion to that in the absence of benzoate anion drastically increased with the dioxane mole fraction and attained a 2500-fold rate acceleration at a dioxane mole fraction of 0.42 (the highest experimentally attainable) when the concentration of benzoate anion was 0.5 M. The proposed mechanism involves proton abstraction by the benzoate anion from the imidazolyl group, followed by proton abstraction by the imidazolyl group from H(2)O, resulting in effective general base-catalysis of hydrolysis. The results of the present paper provide support for the "charge-relay" system in serine proteases.

Benzoates↗
PubMed1977

DTNB modification of SH groups of isocitrate dehydrogenase from Bacillus stearothermophilus purified by affinity chromatography.

A new method of affinity chromatography using blue dextran-Sepharose 4B resin was established to purify NADP+-dependent isocitrate dehydrogenase [EC 1.1.1.42] from Bacillus stearothermophilus in high yield. The purified preparation was found to be homogeneous on disc gel electrophoresis. The SH groups of the enzyme were modified with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) to determine the number of SH groups per molecule and their contribution to the enzyme activity. One SH group was titrated with DTNB per subunit (the native enzyme consisted of two subunits) and after complete denaturation with 4 M guanidine-HCl the number of titratable SH groups remained unchanged. ORD and CD measurements showed that the alpha-helical conformation of the polypeptide backbone was unaffected by DTNB modification, though the near ultraviolet CD spectrum was evidently altered. The fluorescence derived from tryptophanyl residue(s) was quenched by the modification to 30% of the native level, which may indicate the presence of SH in the vicinity of tryptophanyl residue(s). A remarkable decrease of the enzyme activity was detected upon modification with DTNB, but there was some discrepancy between the rate of inactivation and that of modification of SH groups. The presence of substrate and Mg2+ gave partial protection against modification of the SH groups by DTNB. Complete protection of the native enzyme activity against heating at 65 degrees was observed in the presence of substrate and Mg2+, but the thermostability of the enzyme was markedly reduced by modification of the SH groups.

Chromatography, Affinity↗
PubMed1977

Suppression of elevated serum TSH levels in hypothyroidism by fusaric acid.

The effect of Fusaric acid (FA), a specific inhibitor of dopamine beta-hydroxylase, on humna TSH and thyroid hormone concentration (T4 and T3) was evaluated. Healthy subjects showed no significant changes in serum T3,T4 and TSH concentrations following the administration of FA calcium salt (FA-Ca) or placebo. Similarly, administration of FA-Ca for 4 weeks to hypertensive patients failed to produce significant changes in the serum T4 or T3 Resin Sponge Uptake values, and in the TSH and T3 responses to TRH. In contrast, FA-Ca produced a significant reduction on the high basal serum TSH level in patients with primary hypothyroidism. The mean nadir was 25% and ranged from 6 to 61%. As in the case of L-Dopa, the effect of FA-Ca on serum TSH is most clearly demonstrated in patients with primary hypothyroidism. Alterations in brain amines may directly or indirectly suppress pituitary TSH secretion. The possibility of changes in the peripheral distribution or turnover rate of TSH has not been excluded.

Adult↗