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1.
Summary Non-histone chromatin protein (NHCP) fractions were extracted from purified beef thyroid nuclear preparations and tested for the presence of protein kinase activities using several known mediators of thyroid regulation, as well as potential phosphotransferase substrates using purified or partially purified protein kinase activities. The addition of cAMP/3-isobutyl-l-methylxanthine had no effect on NHCP historic kinase activity; the addition of 10 g of the heat-stable cAMP-dependent protein kinase A inhibitor, however, resulted in a 47% reduction in histone H2 kinase activity. Nuclear casein kinase II activity was present in the NHCP fractions as evidenced by the capacity of spermine to stimulate (ED50 = 0.19 mM) and heparin to inhibit (ID50 = 0.09 g/ml) the phosphorylation of casein; further, the phosphotransferase activity could be purified by sequential casein-agarose and spermine-agarose affinity chromatography. Neither calcium-calmodulin nor calcium/phosphatidylserine/diolein had an effect on NHCP casein kinase or histone kinase activities, respectively. The addition of cAMP-dependent protein kinase A catalytic subunit, nuclear casein kinase II, calcium-activated calmodulin-dependent protein kinase and diacylglycerol-activated calcium/phospholipid-dependent protein kinase C activities exhibited distinct phosphorylation patterns when NHCP were used as substrates and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. We conclude that NHCP fraction from beef thyroid: 1) contains both cAMP-dependent protein kinase A catalytic subunit and nuclear casein kinase II and 2) substrates for cAMP-dependent protein kinase A, calcium-activited calmodulin-dependent protein kinase, protein kinase C, and nuclear casein kinase II.Abbreviations NHCP
Non-Histone Chromatin Proteins
- PK-A
cAMP-Dependent Protein Kinase
- CAMPK
Calcium-Activated Calmodulin-Dependent Protein Kinase
- PK-C
Diacylglycerol-Activated Calcium/phospholipid-dependent Protein Kinase
- NK-11
Nuclear Casein Kinase 11
- CK-G
Cytosolic Casein Kinase G or 11
- PMSF
Phenylmethyl Sulfonyl Fluoride
- PKI
the Heat Stable PK-A Inhibitor (Walsh inhibitor)
- SDS-PAGE
Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis
- EDTA
Ethylenediamine Tetraacetic Acid
- EGTA
Ethyleneglycol bis- (B-aminoethyl ether) N,N,N,N,-Tetraacetic Acid
- PS
Phosphatidylserine
- DO
1,2-Diolein 相似文献
2.
B Kerdelhué P Parnet V Lenoir A Schirar F Gaudoux M C Levasseur M Palkovits C Blacker R Scholler 《Journal of steroid biochemistry》1988,30(1-6):161-168
This paper further substantiates the physiological role of beta-endorphin (beta-END) in the control of the cyclic LH secretion and provides new data on the interactions between 17 beta-estradiol (17 beta-E2) and beta-END at both the hypothalamic and pituitary levels. At the hypothalamic level, during the estrous cycle in rats, beta-END concentrations were highest on diestrus I in the arcuate nucleus, median preoptic area and median eminence and lowest at the time of the preovulatory 17 beta-E2 surge on proestrus, before the subsequent preovulatory hypothalamic GnRH and plasma LH surges. Data obtained in ovariectomized 17 beta-E2-treated ewes support the direct involvement of 17 beta-E2 in changes in beta-END and GnRH concentrations in these hypothalamic areas. At the anterior pituitary level, in vitro results obtained using anterior pituitaries from the proestrus morning cycling female rat have shown that 17 beta-E2 strongly suppresses beta-END secretion and that GnRH stimulates the release of beta-END. Furthermore, marked fluctuations were observed for plasma beta-END throughout the menstrual cycle in the woman. Low beta-END concentrations were observed in the period preceding the LH preovulatory surge. Taken together, these results show that: (1) decreases in hypothalamic beta-END concentrations, which are controlled at least by circulating levels of 17 beta-E2, modulate GnRH synthesis and/or release and contribute to the mechanisms which initiate the LH surge; (2) anterior pituitary beta-END might be involved in the mechanisms which terminate the LH surge. 相似文献
3.
Philippe Fragu Colette Brianon Catherine Fourr Jrme Clerc Odile Casiraghi Josette Jeusset Frdrique Omri Sylvain Halpern 《Biology of the cell / under the auspices of the European Cell Biology Organization》1992,74(1):5-18
We attempted to indicate the requirements for biomedical applications of SIMS microscopy. Sample preparation methodology should preserve both the structural and the chemical integrity of the tissue. Furthermore, it is often necessary to correlate ionic and light microscope images. This implies a common methodological approach to sample preparation for both microscopes. The use of low or high mass resolution depends on the elements studied and their concentrations. To improve the acquisition and processing of images, digital imaging systems have to be designed and require both ionic and optical image superimposition. However, the images do not accurately reflect element concentration; a relative quantitative approach is possible by measuring secondary ion beam intensity. Using an internal reference element (carbon) and standard curves the results are expressed in micrograms/mg of tissue. Despite their limited lateral resolution (0.5 microns) the actual SIMS microscopes are very suitable for the resolution of biomedical problems posed by action modes and drug localization in human pathology. SIMS microscopy should provide a new tool for metabolic radiotherapy by facilitating dose evaluation. The advent of high lateral resolution SIMS imaging (less than 0.1 microns) should open up new fields in biomedical investigation. 相似文献
4.
Sabine Lhernould Yannis Karamanos Sylvain Bourgerie Gerard Strecker Raymond Julien Henri Morvan 《Glycoconjugate journal》1992,9(4):191-197
We have previously isolated mannoside and xylomannoside oligosaccharides with one or two terminal reducingN-acetylglucosamine residues from the extracellular medium of white campion (Silene alba) suspension culture. We have now demonstrated the presence of peptide-N
4-(N-acetylglucosaminyl)asparagine amidase (PNGase) activity in cell extracts as well in the culture medium that could explain the production of those compounds. An additional xylomannoside, (GlcNAc)Man3(Xyl)GlcNAc(Fuc)GlcNAc, was characterized, and1H- and13C-NMR assignments for the oligosaccharide Man3(Xyl)GlcNAc(Fuc)GlcNAc were obtained using homonuclear and heteronuclear spectroscopy (COSY).Abbreviations Endo
endo--N-acetylglucosaminidase
- Fuc
fucose
- GlcNAc
N-acetylglucosamine
- Man
mannose
- NMR
nuclear magnetic resonance
- PNGase
peptide-N
4-(N-acetylglucosaminyl)asparagine amidase
- Xyl
xylose 相似文献
5.
Influence of maize root mucilage on soil aggregate stability 总被引:9,自引:0,他引:9
This study was undertaken to determine the effects of root exudates on soil aggregate stability. Root mucilage was collected
from two-month old maize plants (Zea mays L.) Mucilage and glucose solutions were added at a rate of 2.45 g C kg−1 dry soil to silty clay and silt loam soils. Amended soils, placed in serum flasks, were incubated for 42 d with a drying-wetting
cycle after 21 d. Evolved CO2 was measured periodically as well as the water-stable aggregates and soluble sugar and polysaccharide content of the soil.
In mucilage-amended soils CO2 evolution started with a lag phase of 2–3 days, which was not observed in glucose-amended soils. There was then a sharp increase
in evolved CO2 up to day 7. During the second incubation period there were only small differences in evolved C between treatments. Incorporation
of mucilage in both soils resulted in a spectacular and immediate increase in soil aggregate stability. Thereafter, the percent
of water-stable aggregates quickly decreased parallel to microbial degradation. On completion of the incubation, aggregate
stability in the silty clay soil was still significantly higher in the presence of mucilage than in the control. This work
supports the assumption that freshly released mucilage is able to stick very rapidly to soil particles and may protect the
newly formed aggregates against water destruction. On the silty clay, microbial activity contributes to a stabilization of
these established organo-mineral bounds. 相似文献
6.
7.
Integrated cultivation of salmonids and seaweeds in open systems 总被引:2,自引:2,他引:0
Bacterial abundance and production in a vertical profile in Lake Kariba (17dgS), Zimbabwe, were affected by solar irradiance. At the surface, 1.87 × 109 bacteria 1–1 were found and abundance peaked at 10 m (2.5 × 109 bacteria l-1), then decreasing with depth. Bacterial reproduction at the surface(0.145 µg C1–1 h–1) was nearly four times less than the production at 10 m although bacterial numbers were only 26% less. Thus, bacterial production per cell was lower at the surface than deeper down, suggesting that bacterial production is inhibited at the surface.Bacterial production in GF/F filtered lake water in Whirl Pack bags showed an exponential decrease down to 3 m depth. The inhibition was well in accordance with light extinction in the UV region. Phosphatase activity was low in light exposed bags compared to dark, indicating photolysis of extracellular enzymes, or phototransformation of recalcitrant DOM, which substitutes enzyme activity. Hypolimnetic enzyme activity was less affected by solar light than epilimnetic. 相似文献
8.
Cold acclimation and photoinhibition of photosynthesis in Scots pine 总被引:13,自引:0,他引:13
Alla Krivosheeva Da-Li Tao Christina Ottander Gunnar Wingsle Sylvain L. Dube Gunnar Öquist 《Planta》1996,200(3):296-305
Cold acclimation of Scots pine did not affect the susceptibility of photosynthesis to photoinhibition. Cold acclimation did however cause a suppression of the rate of CO2 uptake, and at given light and temperature conditions a larger fraction of the photosystem II reaction centres were closed in cold-acclimated than in nonacclimated pine. Therefore, when assayed at the level of photosystem II reaction centres, i.e. in relation to the degree of photosystem closure, cold acclimation caused a significant increase in resistance to photoinhibition; at given levels of photosystem II closure the resistance to photoinhibition was higher after cold acclimation. This was particularly evident in measurements at 20° C. The amounts and activities of the majority of analyzed active oxygen scavengers were higher after cold acclimation. We suggest that this increase in protective enzymes and compounds, particularly Superoxide dismutase, ascorbate peroxidase, glutathione reductase and ascorbate of the chloroplasts, enables Scots pine to avoid excessive photoinhibition of photosynthesis despite partial suppression of photosynthesis upon cold acclimation. An increased capacity for light-induced de-epoxidation of violaxanthin to zeaxanthin upon cold acclimation may also be of significance.Abbreviations APX
ascorbate peroxidase
- DHA
dehydroascorbate
- DHAR
dehydroascorbate reductase
- Fm
maximal fluorescence when all reaction centres are closed
- Fv/Fm
maximum photochemical yield of PSII
- GR
glutathione reductase
- GSH
reduced glutathione
- Je
rate of photosynthetic electron transport
- MDAR
monodehydroascorbate reductase
- qN
nonphotochemical quenching of fluorescence
- qP
photochemical quenching of fluorescence
- SOD
superoxide dismutase
This work was supported by the Swedish Natural Science Research Council and the National Natural Science Foundation of China. 相似文献
9.
Influence of nitrate on uptake of ammonium by nitrogen-depleted soybean: is the effect located in roots or shoots? 总被引:4,自引:1,他引:3
Saravitz Carole H.; Chaillou Sylvain; Musset Joanne; Raper C. David Jr.; Morot-Gaudry Jean-Francis 《Journal of experimental botany》1994,45(11):1575-1584
In non-nodulated soybean [Glycine max (L.) Merrill cv. Ransom]plants that were subjected to 15 d of nitrogen deprivation inflowing hydroponic culture, concentrations of nitrogen declinedto 1.0 and 1.4mmol Ng1 dry weight in shoots and roots,respectively, and the concentration of soluble amino acids (determinedas primary amines) declined to 40µmol g1 dry weightin both shoots and roots. In one experiment, nitrogen was resuppliedfor 10 d to one set of nitrogen-depleted plants as 1.0 mol m3NH4+ to the whole root system, to a second set as 0.5 mol m3NH4+ plus 0.5 mol m3 NO3 to the whole root system,and to a third set as 1.0 mol m3 NH4+ to one-half ofa split-root system and 1.0 mol m3 NO3 to theother half. In a second experiment, 1.0 mol m3 of nitrogenwas resupplied for 4 d to whole root systems in NH4+ : NO3ratios of 1:0, 9:1, and 1:1. Nutrient solutions were maintainedat pH 6.0. When NH4+ was resupplied in combination with NO3 to thewhole root system in Experiment I, cumulative uptake of NH4+for the 10 d of resupply was about twice as great as when NH4+was resupplied alone. Also, about twice as much NH4+ as NO3was taken up when both ions were resupplied to the whole rootsystem. When NH4+ and NO3 were resupplied to separatehalves of a split-root system, however, cumulative uptake ofNH4+ was about half that of NO3. The uptake of NH4+,which is inhibited in nitrogen-depleted plants, thus is facilitatedby the presence of exogenous NO3, and the stimulatingeffect of NO3 on uptake of NH4+ appears to be confinedto processes within root tissues. In Experiment II, resupplyof nitrogen as both NH4+ and NO3 in a ratio of either1:1 or 9:1 enhanced the uptake of NH4+. The enhancement of NH4+uptake was 1.8-fold greater when the NH4+: NO3-resupplyratio was 1:1 than when it was 9:1; however, only 1.3 timesas much NO3 was taken up by plants resupplied with the1 :1 exogenous ratio. The effect of NO3 on enhancementof uptake of NH4+ apparently involves more than net uptake ofNO3 itself and perhaps entails an effect of NO3uptake on maintenance of K+ availability within the plant. Theconcentration of K+ in plants declined slightly during nitrogendeprivation and continued to decline following resupply of nitrogen.The greatest decline in K+ concentration occurred when nitrogenwas resupplied as NH4+ alone. It is proposed that decreasedavailability of K+ within the NH4+-resup-plied plants inhibitedNH4+ uptake through restricted transfer of amino acids fromthe root symplasm into the xylem. Key words: Ammonium, Glycine max, nitrate, nitrogen-nutrition, nitrogen stress, split-root cultures 相似文献
10.
Mare K. Saba-El-Leil Sylvain Rivard David Morse Mario Cappadocia 《Plant molecular biology》1994,24(4):571-583
A genomic clone of the S11 allele from the self-incompatibility locus (S locus) in Solanum chacoense Bitt. has been isolated by cross-hybridization to the S. chacoense S13 allele and sequenced. The sequence of the S11 allele contains all the features expected for S genes of the Solanaceae, and S11 expression, as assessed by northern blots and RNA-PCR, was similar to that of other S. chacoense S alleles. The S11 protein sequence shares 95% identity with the phenotypically distinct S13 protein of S. chacoense and is the gametophytic S allele with the highest similarity to an existing allele so far discovered. Only 10 amino acid changes differentiate the mature proteins from these two alleles, which sets a new lower limit to the number of changes that can produce an altered S allele specificity. The amino acid substitutions are not clustered, suggesting that an accumulation of random point mutations can generate S allele diversity. The S11 intron is unusual in that it could be translated in frame with the coding sequence, thus suggesting an additional mechanism for the generation of new S alleles. 相似文献