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31.
32.
Does Gibberellic Acid Induce the Transfer of Lipase from Protein Bodies to Lipid Bodies in Barley Aleurone Cells? 总被引:3,自引:0,他引:3 下载免费PDF全文
We have examined the effect of gibberellic acid (GA3) on the distribution of the enzyme responsible for mobilizing storage triacylglycerol in aleurone cells of Hordeum vulgare L. cv Himalaya. Using cellular fractionation techniques, we find that, in cells that have not been exposed to hormone, neutral lipase activity is principally associated with a pellet containing the membranes of protein bodies. If the cells are exposed to GA3 for at least 1 hour, the majority of the lipase activity becomes associated with the lipid body fraction. The nature of the in vivo association between lipid bodies and protein bodies was examined using ultrarapid freezing followed by freeze-fracture electron microscopy. Our analysis indicates that the phospholipid monolayer surrounding the lipid body is directly continuous with the outer leaflet of the bilayer surrounding the protein body. Based on our data, we propose that lipase can be transferred from protein bodies (storage form) to lipid bodies (active form) by lateral diffusion within the plane of the fused phospholipid monolayer, and that the transfer can be controlled by gibberellic acid by an unknown mechanism. 相似文献
33.
Using sucrose density gradient centrifugation in a vertical rotor, we have separated three major binding components contained in hepatic cytosols from C57BL/6 mice and Sprague-Dawley rats. Using this preparative method we have obtained, after a 3-h run of 2.4 ml of crude cytosol from 1,4-bis[2-(3,5-dichlorodipyridyloxy)]benzene-treated C57BL/6 mice (approximately 50 mg of protein: 10,000 fmol of Ah receptor) 50 and 75% yields of isolated Ah receptor and carcinogen-binding protein (4 S binding protein), respectively. Both binding components may be kept at -70 degrees C for several months without loss of activity. A third binding component, which did not sediment in a sucrose density gradient (5-20%), even after a 4-h run at 63,000 rpm, was recovered from the top fractions of gradients. When applied to Sephacryl S-300 columns this component was eluted in the void fraction. Resistant to the direct degradative action of nucleases and proteases, this large complex was sequentially converted to its subcomponents by lipoprotein-lipase, proteinase K, and phospholipases. Only the phospholipases are able to abolish the binding capacity of this light density component (LDC) for [3H]2,3,7,8-tetrachlorodibenzo-p-dioxin: hence, we conclude that phospholipids are the true binders of this radioligand. In vitro, this lipoprotein irreversibly binds many hydrophobic radioligands (2,3,7,8-tetrachlorodibenzo-p-dioxin,3-methylcholanthrene, benzo(a)pyrene, 7,12-dimethylbenz(a)anthracene, and dexamethasone). Using single vertical spin density gradient ultracentrifugation, the major part (80%) of LDC was characterized as a very low-density lipoprotein, and a minor part (20%) as a low-density lipoprotein. This conclusion was supported by the size of LDC particles (about 25-75 nm) observed in electron microscopy. 相似文献
34.
George W. LaBar Jose A. Hernando Casal Carlos Fernandez Delgado 《Environmental Biology of Fishes》1987,19(2):111-117
Synopsis Radio telemetry was used to study the movements of European eels,Anguilla anguilla, in a small (1.2 ha) lake in southwestern Spain in March and April, 1985. Observations were taken on the locations of 7 eels at least once each 2 h for a combined total of 1713 h. The size of individual activity regions varied from 2700 to 1300 m2. Eels covered a larger area at night than during the day, with an average of 23% and 42% of the activity region used during the day and night respectively. Average distance moved between observations was significantly greater at night than in the day. Eels tracked during rainy and cloudy weather were more active during the day and used a larger total area than did those tracked during drier, more stable weather. The standing crop of eels was estimated to be about 77 kg ha–1. 相似文献
35.
C J Fernandez M Lacort J M Gandarias B Ochoa 《Biochemical and biophysical research communications》1987,146(3):1212-1217
Lateral and transversal distribution of cholesterol ester hydrolase activity in rat liver microsomal membranes has been studied. Total cholesterol ester hydrolase activity was found predominantly (75%) in rough microsomes though specific esterase activities were similar in rough and smooth microsomal fractions. The transversal asymmetry of the enzyme was examined using the criteria of protease sensitivity and latency of mannose-6-phosphate phosphatase. Cholesterol ester hydrolase resulted drastically inhibited by proteolysis with trypsin when microsomal integrity had been previously disrupted with sodium deoxycholate or sodium taurocholate. Under these conditions, most lumenal mannose-6-phosphate phosphatase activity was destroyed. However, cholesterol esterase was unaffected by preincubating microsomes with the detergent alone, which led to the complete expression of latent mannose-6-phosphate phosphatase or by preincubating them with trypsin, where less than a 15% of the lumenal mannose-6-phosphate phosphatase was lost. These findings suggest that cholesterol ester hydrolase activity is located on the lumenal surface of the hepatic microsomal vesicles. 相似文献
36.
Catherine Ronin Herman van Halbeek Johannah GM Mutsaers Johannes F G Vliegenthart 《Glycoconjugate journal》1987,4(3):247-254
The lipid-linked precursor ofN-type glycoprotein oligosaccharides was isolated from porcine thyroid microsomes after in cubation with UDP[3H] Glucose. The carbohydrate was released from dolichol pyrophosphate by mild acid hydrolysis, purified by gel filtration and characterized by 500-MHz1H-NMR spectroscopy in combination with enzymatic degradation. The parent oligosaccharide was found to be Glc3Man9Glc-NAc2. The three glucose residues are present in the linear sequence Glcα1-2Glα1-3 Glc, the latter being α(1-3)-linked to one of the mannose residues. In order to establish the branch location of the triglucosyl unit, the parent compound was digested with jack-bean α-mannosidase. The oligosaccharide product was purified by gel filtration, and identified by1H-NMR as Glc3Man5GlcNAc2 lacking the mannose residues A, D2, B and D3. Therefore, the structure of the precursor oligosaccharide is as follows: $$\begin{gathered} c b a D_1 C 4 \hfill \\ Glc\alpha 1 - 2Glc\alpha 1 - 3Glc\alpha 1 - 3Man\alpha 1 - 2Man\alpha 1 - 2Man\alpha 1 \hfill \\ 3 \swarrow 3 2 1 \hfill \\ Man\alpha 1 - 2Man\alpha 1 Man\beta 1 - 4GlcNAc\beta 1 - 4GlcNAc \hfill \\ D_{2 } A 3 6 \hfill \\ Man\alpha 1 \hfill \\ 6 \hfill \\ Man\alpha 1 - 2Man\alpha 1 \nwarrow 4 \hfill \\ D_3 B \hfill \\ \end{gathered} $$ 相似文献
37.
Yéronique Blanquet Catherine Turleau Nicole Créau-Goldberg Chantal Cochet J. de Grouchy 《Human genetics》1987,76(1):102-105
Summary The two probes H3-8 and H2-42, known to be located in 13q14, were mapped by in situ hybridization to either side of the 13 breakpoint of an apparently balanced de novo t(2;13)(p24.3;q14.2) detected in a patient with retinoblastoma as the only phenotypic manifestation. 相似文献
38.
l-Tryptophan decarboxylase (TDC) (EC 4.2.1.27) enzyme activity was induced in cell suspension cultures of Catharanthus roseus after treatment with a Pythium aphanidermatum elicitor preparation. The enzyme was extracted from lyophilized cells containing high levels of TDC and the protein was purified to homogeneity. The pure protein was used to produce highly specific polyclonal antibodies, and an enzyme-linked immunosorbent assay (ELISA) was developed to quantitate the level of TDC antigen during seedling development and in leaves of the mature plant. Western immunoblotting of proteins after SDS-PAGE with anti-TDC antibodies detected several immunoreactive proteins (40, 44, 54.8, 55, and 67 kilodaltons) which appeared at different stages during seedling development and in leaves of the mature plant. The major 54.8 and 55 kilodalton antigenic proteins in immunoblots appeared transiently between days 1 to 5 and 5 to 8 of seedling development, respectively. The 54.8 kilodalton protein was devoid of TDC enzyme activity, whereas the appearance of the 55 kilodalton protein coincided with the appearance of this decarboxylase activity. The minor immunoreactive proteins (40, 44, and 67 kilodaltons) appeared after day 5 of seedling development and in older leaves of the mature plant, and their relationship, if any, to TDC is presently unknown. Results suggest that the synthesis and degradation of TDC protein is highly regulated in Catharanthus roseus and that this regulation follows a preset developmental program. 相似文献
39.
Catherine Digonnet-Kerhoas Gilles Gay Jean Claude Duplan Christian Dumas 《Planta》1989,179(2):165-170
During ageing of the short-lived pollen grains of Cucurbita pepo L., water loss was examined in relation to viability using biophysical (1H-nuclear magnetic resonance, NMR) and cytological methods (fluorochromatic reaction test, freezefracture and scanning electron microscopy). A semi-logarithmic representation of the pollen weight loss demonstrated the complexity of the dehydration process. A the study of proton loss using 1H-NMR indicated that two major releases water of had taken place, each with different flux rates. Pulse 1H-NMR experiments showed the occurrene of non-exponential signal decay as a function of time, indicating the existence of different fractions of water in a pollen grain sample. These fractions leave the pollen grain at different times during pollen dehydration, and one of them (that of the so-called vital water) can be related to pollen viability. The quantity of protons giving a signal during pulse 1H-NMR experiments was very low when the pollen grains were judged to be dead according to the fluorochromatic test. Freeze-fracture replicas of these dead pollen grains (less than 25% water content) showed that the plasma membrane had become detached from the intine surface; this ultrastructural feature might therefore be involved in the loss of pollen viability.Abbreviations A
initial amplitude of the NMR signal
- A2
quantity of water charcterized by T2-2
- A5
quantity of water characterized by T2–5
- FCR
fluorochromatic reaction
- NMR
nuclear magnetic resonance
- T2
transverse relaxation time
- T2-2
T2 measured with 2 ms between each pulse of radiofrequency
- T2–5
T2 measured with 5 ms between each pulse of radiofrequency 相似文献
40.
When following the pattern of the disappearance of NH
4
+
–N from ammonium sulfate applied to the flooded soil-rice plant system (field and greenhouse experiments) during a growing season, it was observed that the lowest NH
4
+
–N level coincided with the highest value of NR activity in the leaves. Nitrate was detected in both the root and shoot systems of the rice plants and autotrophic nitrifiers (Nitrosomonas and Nitrobacter) were particularly abundant. Since it was also demonstrated in this work that the NR activity of rice plants grown with nitrate fertilization (growth chamber culture experiments) was inducible by its substrate, it can be assumed that NH
4
+
–N oxidation takes place in the water-logged soil studied. Therefore, the occurrence of the nitrification process following NH
4
+
–N fertilizer application can be predicted by thein vitro orin situ evaluation of the NR activity of the rice leaf as an indicator. 相似文献