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1.
The subcellular distribution of ATP, ADP, creatine phosphate and creatine has been analyzed by fast detergent fractionation of isolated frog heart cells. Digitonin fractionation (0.5 mg/ml, 10 s at 2 degrees C in 20 mM 4-morpholinepropanesulfonic acid/3 mM EDTA/230 mM mannitol medium) was used to separate mitochondria and myofilaments from cytosol. To separate myofilaments from the other cellular compartments. Triton X-100 was used (2%, 15 s in the same medium as digitonin). For either resting or beating cells the total cellular contents of ATP, ADP, creatine phosphate and creatine was similar, nevertheless the O2 consumption was 6-times higher. The compartmentation of these metabolites was also identical. Myofilaments contain 1.1 nmol ADP per mg total cellular proteins. In the cytosolic compartment the metabolite concentrations, all measured in nmol per mg total cellular proteins, were: ATP, 13; ADP, 0.25-0.05; creatine phosphate, 18.5 and creatine, 14. This indicated that the reaction catalyzed by creatine kinase was in a state of (or near) equilibrium. 相似文献
2.
Rajewsky B. Nay U. Schmidt-Burbach G. Friedrich E. 《Radiation and environmental biophysics》1967,4(2):183-186
Zusammenfassung Es wurden künstliche radioaktive Teilchen in Form von im Durchmesser 5–20 m großen Plexiglasperlen hergestellt, in die eine organische Scandiumverbindung homogen eingelagert ist. Nach eintägiger Aktivierung der Perlen im Reaktor kann bei einem Teilchen von 10 m Durchmesser eine Scandiumaktivität bis zu 10–10 Ci erreicht werden. 相似文献
3.
M. A. Lelu K. Klimaszewska G. Pflaum C. Bastien 《In vitro cellular & developmental biology. Plant》1995,31(1):15-20
Summary Cotyledonary somatic embryos ofLarix × leptoeuropaea that developed after various maturation times on media containing abscisic acid showed different frequencies of conversion
into plants. Drying of these somatic embryos under high relative humidity (RH) before germination improved plantlet recovery
and eliminated differences in the performance of somatic embryos matured for different times. However, dehydration of somatic
embryos under 98% RH to a water content below that of zygotic embryos excised from mature seeds (0.97 and 1.36 g H2O/g dry weight, respectively) showed a strong positive correlation between longer maturation time and desiccation tolerance.
Drying somatic embryos at 4° C under 59% RH for 1 wk resulted in desiccation to a water content of 0.30 g H2O/g dry weight, which was the closest to the hydration state of zygotic embryos in dried, stored seeds (0.20 g H2O/g dry weight). Under this condition, only somatic embryos matured for 5 wk germinated and produced plantlets at a relatively
high frequency (73 and 41%, respectively). 相似文献
4.
Primary sequence of the glucanase gene from Oerskovia xanthineolytica. Expression and purification of the enzyme from Escherichia coli 总被引:14,自引:0,他引:14
S H Shen P Chrétien L Bastien S N Slilaty 《The Journal of biological chemistry》1991,266(2):1058-1063
A 2.7-kilobase fragment of DNA from Oerskovia xanthineolytica containing the gene for a beta-1,3-glucanase has been isolated and its complete nucleotide sequence determined. The sequence was found to contain two large open reading frames. Purification of the mature native enzyme and subsequent amino-terminal sequencing defined the glucanase gene in one reading frame which potentially encodes a protein of 548 amino acids. We have expressed this glucanase gene in Escherichia coli under control of the lacUV5 promoter and found the product to be secreted into the periplasm as a mature enzyme of about the same molecular weight as that of the native protein. The recombinant enzyme was purified to near homogeneity by a single step of high performance liquid chromatography. The ability of the recombinant enzyme to digest beta-glucan substrates and to lyse viable yeast cells was found to be indistinguishable from that of the native protein. Deletion of the cysteine-rich carboxyl-terminal 117 amino acids of the enzyme, which also contain two duplicated segments, abolished the lytic activity but did not significantly affect the glucanase function of the protein. The possible involvement of this domain in interaction with the yeast cell wall is discussed. 相似文献
5.
The authors have studied the in vitro conversion of 18 hydroxycorticostérone to aldosterone (18 oxidation) by duck adrenal subcellular fractions. Considering the new hypothesis about the mechanism of this step (hydroxylation mechanism) the authors have investigated a possible relationship between this reaction and cytochrome P450. With experimental conditions described, data show that metyrapone, a cytochrome P450 competitive inhibitor does not inhibit 18 oxidation. In contrast, 18 oxidation is inhibited by spirolactones (spironolactones, canrenone, potassium canrenoate). These compounds act at the cytochrome P450 level but have also an uncoupling effect which has been recently discovered. The effects of metyrapone and spirolactones on 18 oxidation as well as the different behaviour between biologicaly and organically synthetised 18 hydroxycorticosterone allow us to propose hypotheses for the mechanism of this step. 相似文献
6.
Victoire Cardot‐Ruffino Vronique Chauvet Cassandre Caligaris Adrien Bertrand‐Chapel Nicolas Chuvin Roxane M. Pommier Ulrich Valcourt David Vincent Sylvie Martel Sophie Aires Bastien Kaniewski Pierre Dubus Philippe Cassier Stphanie Sentis Laurent Bartholin 《Genesis (New York, N.Y. : 2000)》2020,58(5)
Recombination systems represent a major breakthrough in the field of genetic model engineering. The Flp recombinases (Flp, Flpe, and Flpo) bind and cleave DNA Frt sites. We created a transgenic mouse strain ([Fsp1‐Flpo]) expressing the Flpo recombinase in fibroblasts. This strain was obtained by random insertion inside mouse zygotes after pronuclear injection. Flpo expression was placed under the control of the promoter of Fsp1 (fibroblast‐specific protein 1) gene, whose expression starts after gastrulation at Day 8.5 in cells of mesenchymal origin. We verified the correct expression and function of the Flpo enzyme by several ex vivo and in vivo approaches. The [Fsp1‐Flpo] strain represents a genuine tool to further target the recombination of transgenes with Frt sites specifically in cells of mesenchymal origin or with a fibroblastic phenotype. 相似文献
7.
8.
Roque d’orbcastel Emmanuelle Bettarel Yvan Dellinger Marion Sadoul Bastien Bouvier Thierry Amandé Justin Monin Dagorn Laurent Geffroy Benjamin 《Environmental Biology of Fishes》2021,104(6):725-732
Environmental Biology of Fishes - Cortisol is recognized as a physiological indicator of stress in fish. However, this hormone is typically measured in plasma samples. In this study, cortisol... 相似文献
9.
Bastien Christ Iris Süssenbacher Simone Moser Nicole Bichsel Aurelie Egert Thomas Müller Bernhard Kr?utler Stefan H?rtensteiner 《The Plant cell》2013,25(5):1868-1880
Nonfluorescent chlorophyll catabolites (NCCs) were described as products of chlorophyll breakdown in Arabidopsis thaliana. NCCs are formyloxobilin-type catabolites derived from chlorophyll by oxygenolytic opening of the chlorin macrocycle. These linear tetrapyrroles are generated from their fluorescent chlorophyll catabolite (FCC) precursors by a nonenzymatic isomerization inside the vacuole of senescing cells. Here, we identified a group of distinct dioxobilin-type chlorophyll catabolites (DCCs) as the major breakdown products in wild-type Arabidopsis, representing more than 90% of the chlorophyll of green leaves. The molecular constitution of the most abundant nonfluorescent DCC (NDCC), At-NDCC-1, was determined. We further identified cytochrome P450 monooxygenase CYP89A9 as being responsible for NDCC accumulation in wild-type Arabidopsis; cyp89a9 mutants that are deficient in CYP89A9 function were devoid of NDCCs but accumulated proportionally higher amounts of NCCs. CYP89A9 localized outside the chloroplasts, implying that FCCs occurring in the cytosol might be its natural substrate. Using recombinant CYP89A9, we confirm FCC specificity and show that fluorescent DCCs are the products of the CYP89A9 reaction. Fluorescent DCCs, formed by this enzyme, isomerize to the respective NDCCs in weakly acidic medium, as found in vacuoles. We conclude that CYP89A9 is involved in the formation of dioxobilin-type catabolites of chlorophyll in Arabidopsis. 相似文献
10.
Hélène Delatte Céline Toty Sébastien Boyer Anthony Bouetard Fanny Bastien Didier Fontenille 《PLoS neglected tropical diseases》2013,7(3)
Arbovirus vector dynamics and spread are influenced by climatic, environmental and geographic factors. Major Chikungunya and Dengue fever outbreaks occurring the last 10 years have coincided with the expansion of the mosquito vector Aedes albopictus to nearly all the continents. We characterized the ecological (larval development sites, population dynamics, insemination and daily survival rates) and genetic (diversity, gene flow, population structure) features of two Aedes albopictus populations from distinct environments (rural and urban) on Réunion Island, in the South-West Indian Ocean. Microsatellite analysis suggests population sub-structuring Ae. albopictus populations. Two genetic clusters were identified that were significantly linked to natural versus urban habitats with a mixed population in both areas. Ae. albopictus individuals prefer urban areas for mating and immature development, where hosts and containers that serve as larval development sites are readily available and support high population densities, whereas natural environments appear to serve as reservoirs for the mosquito. 相似文献