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11.
Analysis of products formed in Chlorella vulgaris 11 h cellsduring photosynthesis in air containing 3,000 ppm 14CO2 at varioustemperatures revealed that the level of 14C-starch was maximumaround 2024?C and decreased with further rise in temperatureuntil 40?C, while 14C-sucrose greatly increased at temperaturesabove about 28?C. Elevating the temperature from 20 to 38?Cduring photosynthetic 14CO2 fixation resulted in a remarkabledecrease in 14C in starch and a concomitant increase in 14Cin sucrose. This conversion of starch to sucrose when shiftingthe temperature from 20 to 38?C proceeded even in the dark.Hydrolysis of sucrose by rß-fructosidase showed that,irrespective of the experimental conditions, the radioactivitiesin sucrose were equally distributed between glucose and fructose.The enhancement of starch degradation with temperature risewas more remarkable than that of the activity of ribulose bisphosphatecarboxylase from the same cells. When Chlorella cells whichhad been preloaded with 14C-starch after photosynthesis for30 min at 20?C were incubated in the dark for an additional30 min at 20?C, 14C-starch was degraded by only about 4%. However,the values after 30-min dark incubation at 28, 32, 36 and 40?Cwere increased by about 10, 19, 36 and 50%, respectively. Duringthe temperature-dependent conversion of starch to sucrose, nosignificant amount of radioactivity accumulated in free glucoseand maltose. (Received October 27, 1981; Accepted January 9, 1982) 相似文献
12.
Chlorella vulgaris 11h cells grown in air enriched with 4% CO2(high-CO2 cells) had carbonic anhydrase (CA) activity whichwas 20 to 90 times lower than that of algal cells grown in ordinaryair (containing 0.04% CO2, low-CO2 cells). The CO2 concentrationduring growth did not affect either ribulose 1,5-bisphosphate(RuBP) carboxylase activity or its Km for CO2. When high-CO2 cells were transferred to low CO2 conditions,CA activity increased without a lag period, and this increasewas accompanied by an increase in the rate of photosynthetic14CO2 fixation under 14CO2-limiting conditions. On the otherhand, CA activity as well as the rate of photosynthetic 14CO2fixation at low 14CO2 concentrations decreased when low-CO2cells were transferred to high CO2 conditions. Diamox, an inhibitor of CA, at 0.1 mM did not affect photosynthesisof low-CO2 cells at high CO2 concentration (0.5%). Diamox inhibitedphotosynthesis only under low CO2 concentrations, and the lowerthe CO2 concentration, the greater was the inhibition. Consequently,the CO2 concentration at which the rate of photosynthesis attainedone-half its maximum rate (Km) greatly increased in the presenceof this inhibitor. When CO2 concentration was higher than 1%, the photosyntheticrate in low-CO2 cells decreased, while that in high-CO2 cellsincreased. Fractionation of the low-CO2 cells in non-aqueous medium bydensity showed that CA was fractionated in a manner similarto the distribution of chlorophyll and RuBP carboxylase. These observations indicate that CA enhances photosynthesisunder CO2-limiting conditions, but inhibits it at CO2 concentrationshigher than a certain level. The mechanism underlying the aboveregulatory functions of CA is discussed.
1This work was reported at the International Symposium on PhotosyntheticCO2-Assimilation and Photorespiration, Sofia, August, 1977 (18).Requests for reprints should be addressed to S. Miyachi, RadioisotopeCentre, University of Tokyo, Bunkyo-ku, Tokyo 113, Japan. (Received December 11, 1978; ) 相似文献
13.
14.
M Tozuka H Hidaka M Miyachi K Furihata T Katsuyama M Kanai 《Biochimica et biophysica acta》1992,1165(1):61-67
A new apolipoprotein complex designated as the apo(AII-E2-AII) complex was identified in the lipoprotein fractions of human plasma with apoE phenotypes containing apoE2 (E4/E2, E3/E2, and E2/E2). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by an immunoblotting assay using anti-apoE or anti-apoAII antibodies, established that the apo(AII-E2-AII) complex, with a molecular weight of 58,000, was identical to the complex consisting of apoE and apoAII, and that it also dissociated following reduction with beta-mercaptoethanol. This new complex was also demonstrated to be distinct from the apo(E-AII) complex and apoE monomer by isoelectric focusing, in the samples that were not treated with beta-mercaptoethanol. In apoE phenotype E3/E2, the apo(AII-E2-AII) complex was primarily included in the high-density lipoprotein (HDL, 1.063 < d < 1.21 g/ml) fraction, but was also observed in a small quantity in the very-low-density lipoprotein (VLDL, d < 1.006 g/ml) fraction. For further characterization, the apo(AII-E2-AII) complex was isolated by preparative SDS-PAGE, and no contamination of apo(E-AII) complex and apoE monomer was detected by immunoblotting assay using an anti-apoE antibody. It was confirmed by an enzyme-linked immunosorbent assay (ELISA) system that a molecular ratio between apoAII monomer and apoE in the isolated apo(AII-E2-AII) complex was approx. 2, when the apo(E-AII) complex was used as a standard with the ratio of 1:1. It indicates that the apo(AII-E2-AII) complex is formed from two molecules of apoAII monomer and one molecule of apoE.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
15.
T Kamo K Shimogawara H Fukuzawa S Muto S Miyachi 《European journal of biochemistry》1990,192(2):557-562
Carbonic anhydrase purified from the cell surface of Chlamydomonas reinhardtii was inactivated by treatment with dithiothreitol. This treatment caused dissociation of the holoenzyme into 35-kDa (A) and 4-kDa (B) subunits as revealed by SDS/PAGE. The 35-kDa subunit was further separated into two components A1 (35 kDa) and A2 (36.5 kDa) by SDS/PAGE using a gradient gel. These two components have the same amino acid sequence up to at least the 10th amino acid from the N-terminus. The molecular masses were estimated at 76 kDa and 35 kDa for the holoenzyme and the large subunit, respectively, and the molar ratio of the former to the latter at 1:2, by using the techniques of low-angle laser light-scattering photometry and precision differential refractometry combined with gel-filtration HPLC. The molar ratio of the 35-kDa/4-kDa subunits was estimated at 1:1 the gel-filtration HPLC monitored with precision differential refractometry. Atomic-absorption spectrophotometry revealed that the holoenzyme contains two atoms of zinc. These results suggest that the holoenzyme is a heterotetramer composed of two large subunits (A1 and A2) and two small subunits (B). 相似文献
16.
In this study we examined the expression of EGF receptor mRNA after EGF administration in hCG producing tumor (choriocarcinoma). We transplanted the tissue of choriocarcinoma into female nude mice and investigated the effects of EGF on the growth of tumors, the binding activity of EGF receptor and the expression of EGF receptor mRNA in the tumor tissues. Two doses of EGF 5.0 micrograms, 50 micrograms and phosphate buffered saline as a control were injected subcutaneously every day for four weeks. Removed tumors were used for immunocytochemical studies and EGF receptor mRNA investigations. HCG and EGF receptors were detected immunocytochemically in the tumor. The low dose EGF employed stimulated the tumor growth while the high dose EGF inhibited the tumor growth compared with that of the control group. The binding activity of EGF receptor and the expression of EGF receptor mRNA also changed in accordance with the stimulation or inhibition of tumor growth. The growth of hCG producing tumor by EGF administration appeared to be dependent upon the binding activity of EGF receptor and the expression of EGF receptor mRNA. 相似文献
17.
Akio Shiraishi Nobuyuki Miyachi 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1976,110(1):97-109
Summary The proboscis extension reponse of the blowfly during stimulation of the tarsal sugar receptors was inhibited by the presence of NaCl. Acceptance thresholds for sucrose in various concentrations of NaCl were measured. The median acceptance thresholds for sucrose in mixtures of 0.01, 0.25, 0.5 and 1.0 M NaCl were 1.8 × 10–3, 6.0 × 10–3, 1.2 × 10–2, and 2.0 × 10–2 M, respectively. Concentration-response curves for sucrose in the tarsal D-type sugar receptor shifted to the right under the existence of high concentration of NaCl. Number of impulses per D-type sugar receptor at the median acceptance thresholds described above were 7.5, 8.4, 6.8 and 10.4 for the first 0.1 s of stimulation, respectively. The average number was 8.2 impulses per 0.1 s. Comparisons were made between the behavioral acceptance thresholds (1) on one leg exposed to sucrose mixed with 0.01 M NaCl and (2) on two contralateral legs, one of which was exposed to sucrose in 0.01 M NaCl and the other to 0.5 M NaCl alone. The acceptance thresholds from two experiments agreed with each other. The median threshold value was 1.7 × 10–2 M sucrose. Behavioral inhibition by NaCl in mixtures with sucrose can be explained by its peripheral inhibition of sugar receptors.This research was supported in part by ITO foundation and Scientific Research Fund from the Ministry of Education of Japan. 相似文献
18.
In order to examine the hypothesis that the thyroidal responsiveness to TSH is under the influence of thyroid hormone, following the T3 injection to the mice, serum T3 concentrations and the response of thyroid tissue to a fixed dose of TSH in terms of intracellular colloid droplet formation was studied. The colloid droplets induced by TSH was significantly reduced when serum T3 was decreasing, while it was significantly increased when serum T3 was increasing. This results demonstrate for the first time the existence of short feed-back loop regulating intra-thyroidal function by circulating T3. To delineate the possible mechanism of action of T3, the thyroid gland of mouse whose serum T3 concentration was elevated by injecting 50 mug T3, was incubated with TSH in vitro. TSH-induced cyclic AMP generation was not inhibited at all but colloid droplet formation was significantly inhibited in the thyroid tissue of the animal whose serum T3 concentration was enormously high. Thus, it was demonstrated that the site at which T3 affects is beyond cylcic AMP generation but prior to endocytosis, being consistent with our previous results. 相似文献
19.
A sensitive radioimmunoassay for plasma betamethasone 17-benzoate has been developed. The antiserum used was obtained by immunizing rabbits with betamethasone 17-benzoate-21-hemisuccinate-bovine-serum-albumin conjugate. All of the endogenous steroids tested cross reacted less than 0.10%. A standard curve was established with a useful range from 0.05-5 ng. Reliability criteria were satisfactory. Measurement of plasma concentrations of betamethasone 17-benzoate was performed in patients and in rabbits following occlusive dressing of betamethasone 17-benzoate cream and gel base. 相似文献
20.
Yu Ono Kenta Katayama Tomoki Onuma Kento Kubo Hayato Tsuyuzaki Michiaki Hamada Masamitsu Sato 《Nucleic acids research》2022,50(19):11229
Non-coding RNAs (ncRNAs) ubiquitously exist in normal and cancer cells. Despite their prevalent distribution, the functions of most long ncRNAs remain uncharacterized. The fission yeast Schizosaccharomyces pombe expresses >1800 ncRNAs annotated to date, but most unconventional ncRNAs (excluding tRNA, rRNA, snRNA and snoRNA) remain uncharacterized. To discover the functional ncRNAs, here we performed a combinatory screening of computational and biological tests. First, all S. pombe ncRNAs were screened in silico for those showing conservation in sequence as well as in secondary structure with ncRNAs in closely related species. Almost a half of the 151 selected conserved ncRNA genes were uncharacterized. Twelve ncRNA genes that did not overlap with protein-coding sequences were next chosen for biological screening that examines defects in growth or sexual differentiation, as well as sensitivities to drugs and stresses. Finally, we highlighted an ncRNA transcribed from SPNCRNA.1669, which inhibited untimely initiation of sexual differentiation. A domain that was predicted as conserved secondary structure by the computational operations was essential for the ncRNA to function. Thus, this study demonstrates that in silico selection focusing on conservation of the secondary structure over species is a powerful method to pinpoint novel functional ncRNAs. 相似文献