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排序方式: 共有298条查询结果,搜索用时 15 毫秒
1.
K Yoshida T Sakurada H Kitaoka H Fukazawa N Kaise K Kaise M Yamamoto S Saito K Yoshinaga 《Endocrinologia japonica》1983,30(2):211-217
Monodeiodination of T4 to T3 and rT3 in the intact cells of dog renal tubuli and glomeruli was investigated. The tubuli and glomeruli were obtained by a sieve method. T4 (2 micrograms/ml) was incubated in Tris-HCl buffer, pH 7.5, with renal cells (180 micrograms protein/ml) and 5 mM DTT for 1 h at 37 degrees C and the T3 and rT3 generated during incubation were measured by specific radioimmunoassays. In order of decreasing activity, dog renal cortical tubuli, cortical homogenate, glomeruli and medullary tubuli were capable of converting T4 to T3. Net rT3 production from T4 in cortical tubuli was also greater than that in cortical homogenate. The conversion of T4 to T3 and also to rT3 in cortical tubuli was enzymatic in nature, since the reactions showed dependence on time and protein concentration; instability to heating; temperature and pH optimum. The production of T3 and rT3 from T4 was maximum at pH 6.5 and at pH 9.5, respectively, indicating that two different enzymic systems, a 5- and a 5'-monodeiodinase, might be involved in the deiodination of the tyrosyl and the phenolic ring of T4 in dog kidney. 相似文献
2.
Characterization and physiological function of glutathione reductase in Euglena gracilis z. 总被引:2,自引:0,他引:2
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The purified glutathione reductase was homogeneous on polyacrylamide-gel electrophoresis. It had an Mr of 79,000 and consisted of two subunits with a Mr of 40,000. The activity was maximum at pH 8.2 and 52 degrees C. It was specific for NADPH but not for NADH as the electron donor; the reverse reaction was not observed. The Km values for NADPH and GSSG were 14 and 55 microM respectively. The enzyme activity was markedly inhibited by thiol inhibitors and metal ions such as Hg2+, Cu2+ and Zn2+. Euglena cells contained total glutathione at millimolar concentration. GSH constituted more than 80% of total glutathione in Euglena under various growth conditions. Glutathione reductase was located solely in cytosol, as were L-ascorbate peroxidase and dehydroascorbate reductase, which constitute the oxidation-reduction cycle of L-ascorbate [Shigeoka et al. (1980) Biochem. J. 186, 377-380]. These results indicate that glutathione reductase functions to maintain glutathione in the reduced form and to accelerate the oxidation-reduction of L-ascorbate, which scavenges peroxides generated in Euglena cells. 相似文献
3.
The pellicle was isolated from the cell homogenate obtained on sonication of Euglena gracilis z grown aerobically under illumination and purified by a combination of differential and sucrose density gradient centrifugations. The purity and homogeneity of the pellicle fragments were determined by an electron microscopic method and biochemical analysis of the components. The protein, lipid, and sugar contents of the purified pellicle were 68.7, 17.9, and 13.5%, respectively. The equilibrium density of pellicle fragments was 1.21 g/cm3. SDS-polyacrylamide gel electrophoresis revealed that the pellicle contained 50 mol% of nonpolar amino acids. The constituents of the lipid and sugar were very different from those of the cell membrane of other organisms. 相似文献
4.
Euglena aquacobalamin reductase (NADPH: EC 1.6.99.-) was purified, and its subcellular distribution was studied to elucidate the mechanism of the conversion of hydroxocobalamin to 5'-deoxyadenosylcobalamin. The enzyme was found in the mitochondria. It was purified about 150-fold over the Euglena mitochondrial extract in a yield of 38%. The purified enzyme was homogeneous in polyacrylamide gel electrophoresis. Spectra of the purified enzyme showed that it was a flavoprotein. The molecular weight of the enzyme was calculated to be 66,000 by Sephadex G-100 gel filtration and 65,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was specific to NADPH with an apparent Km of 43 microM and to hydroxocobalamin with an apparent Km of 55 microM. The enzyme did not require FAD or FMN as a cofactor. The optimum pH and temperature were 7.0 and 40 degrees C, respectively. 相似文献
5.
Euglena contains glutamate:glyoxylate aminotransferase (GGT) both in mitochondria and in cytosol. Both isoforms were separated from each other by DEAE-cellulose chromatography. The mitochondrial enzyme had an apparent Km of 1.9 mM for glutamate and the cytosolic enzyme 52.6 mM. Mitochondrial GGT was further purified by ammonium sulfate fractionation, isoelectric focusing, and gel chromatography. It had a molecular weight of 141,000 and an isoelectric point of pH 4.88; the optimum pH was 8.5. Its apparent Km values for glutamate and for glyoxylate were 2.0 and 0.25 mM, respectively. In addition to glutamate, mitochondrial GGT used 5-hydroxytryptophan, tryptophan, and cysteine as amino donors in the transamination to glyoxylate. Alanine did not support the activity. The relative activity of the enzyme for amino acceptors on the transamination from glutamate was 4-hydroxyphenylpyruvate greater than phenylpyruvate greater than glyoxylate greater than hydroxypyruvate. Pyruvate and 2-oxoglutarate were not used in the reaction. Evidence that GGT functions mainly in the irreversible transamination between glutamate and glyoxylate is presented. The functional significance of GGT in the glycolate pathway of Euglena is also discussed. 相似文献
6.
In Euglena gracilis a malonyl-CoA-independent fatty acid-synthetic system, in which fatty acids are synthesized directly from acetyl-CoA as both primer and C2 donor, occurs in mitochondria, and the system contributes to the wax ester fermentation. The activity of fatty acid synthesis in the mitochondrial system was enhanced about six times when an artificial acetyl-CoA-regenerating system was present, indicating that the fatty acid-synthetic activity is controlled by the ratio of acetyl-CoA against CoA. When fatty acids were synthesized using pyruvate instead of acetyl-CoA as substrate, a high activity, about 30 times higher than that from acetyl-CoA, was found under anaerobic conditions (below 10(-5) M oxygen), while in aerobiosis fatty acids were not synthesized at all. CoA, NADH, and NADP+ were required as cofactors for fatty acid synthesis from pyruvate. It was indicated that high activity of fatty acid synthesis from pyruvate due to the high ratio of acetyl-CoA against CoA was maintained by the action of the oxygen-sensitive pyruvate dehydrogenase found in Euglena mitochondria. When [2-14C]pyruvate was fed into intact mitochondria under anaerobic conditions, radioactive fatty acids were formed in the presence of malate, which provided reducing power for the matrix. 相似文献
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9.
The present study was conducted to determine the cell-cycle dependency of various actions of IGF-I in Balb/c 3T3 cells. When autophosphorylation of the IGF-I receptor was determined in [32P]-labelled cells, IGF-I increased radioactivity in a 100 K-Da phosphoprotein, presumably beta-subunit of the IGF-I receptor, both in quiescent and in primed competent cells. Likewise, IGF-I stimulated uptake of [3H]deoxyglucose independent of the cell cycle. In contrast, IGF-I increased calcium entry, radioactivity in [3H]diacylglycerol, and [3H]thymidine incorporation in primed competent cells while these reactions were not induced by IGF-I in quiescent cells. The latter three reactions were attenuated when cells were pretreated with pertussis toxin. These results indicate that some, but not all, of the actions of IGF-I are dependent on the cell cycle in Balb/c 3T3 cells. They also suggest that a pertussis-toxin-sensitive G protein may be involved in the cell-cycle-dependent actions of IGF-I. 相似文献
10.
K Takano N Yamashita I Kojima M Kitaoka A Teramoto E Ogata 《Biochemical and biophysical research communications》1992,182(3):1408-1415
Activin A stimulated synthesis and secretion of intact FSH in dispersed human FSH-secreting adenoma cells. Significant stimulation was observed after 24 hr. Activin A caused an increase in Ca2+ concentration ([Ca2+]i). This response occurred soon after the activin A action. These effects were blocked in Ca(2+)-deficient medium and by nitrendipine (5 microM). Somatostatin inhibited the activin A-induced intact FSH secretion and the [Ca2+]i response. These findings indicated that Ca2+ influx through voltage-gated Ca2+ channel was involved in the activin A induced synthesis and secretion of intact FSH. 相似文献