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1.
Ivor P. B. Tittawella 《Molecular & general genetics : MGG》1976,145(2):223-226
Summary By using rifampicin to increase the rate of and synthesis in a heterodiploid strain of E. coli carrying the mutation rif
pr (Km7), which codes for a rifampicin sensitive RNA polymerase to which the drug binds weakly, and the dominant mutation rif
D
R
, which codes for a rifampicin resistant RNA polymerase, the concentration of these subunits in the cell was increased 1.6 fold. Measurements made after removal of rifampicin from the cells showed that the excess and subunits did not reduce the rate of their own synthesis below normal. 相似文献
2.
The role of de novo synthesis in the regulation of adenosine 5-phosphosulfate sulfotransferase activity by H2S inLemna minor L. was investigate using density labeling with15N applied as15NO
3
–
in the culture medium. While adenosine 5-phosphosulfate sulfotransferase activity was rapidly reduced by H2S and rapidly recovered upon removal of H2S, O-acetyl-L-serine sulfhydrylase (EC 4.2.99.8) did not show changes in extractable activity in response to H2S and could therefore be used as an internal marker enzyme for density labeling. The incorporation of15N into adenosine 5-phosphosulfate sulfotransferase was strongly reduced upon transfer of plants into a H2S-containing atmosphere. Half-maximal labeling was reached only after 70–80 h compared to 40–50 h in the control. After removal of H2S, adenosine 5-phosphosulfate sulfotransferase activity increased to the initial level within 20 h, and the enzyme reached halfmaximal labeling after only 15 h. The time course of the density increase of O-acetyl-L-serine sulfhydrylase was not affected very significantly by H2S. These results provide evidence that de novo synthesis of enzyme protein is involved in the regulation of adenosine 5-phosphosulfate sulfotransferase activity by H2S.Abbreviations APS
adenosine 5-phosphosulfate
- APSSTase
adenosine 5-phosphosulfate sulfotransferase
- BSA
Bovine serum albumine
- DTE
dithioerythritol
- OAS
O-acetyl-L-serine
- OASSase
O-acetyl-L-serine sulfhydrylase
- POPOP
1,4-bis-(5-phenyl-2-oxazolyl)-benzene
- PPO
2,5-diphenyloxazole
This is no. 9 in the series Regulation of Sulfate Assimilation in Plants 相似文献
3.
M. Rita I. Young Gayle McCloskey Mark A. Wright Annette Schmidt Pak 《Cancer immunology, immunotherapy : CII》1994,38(1):9-15
By secreting granulocyte/macrophage colonystimulating factor (GM-CSF), metastatic Lewis lung carcinoma (LLC-LN7) tumors induce the appearance of myelopoiesis-associated immune-suppressor cells that resemble granulocytic-macrophage (GM) progenitor cells. The presence of these GM-suppressor cells in mice bearing LLC-LN7 tumors was associated with a reduced capacity of splenic T cells to proliferate in response to interleukin-2 (IL-2). Administration of low doses of 100 U interferon (IFN) plus 10 U tumor necrosis factor (TNF) to the tumor bearers, a combination treatment that we previously showed to diminish the presence of GM-suppressor cells synergistically, restored proliferative responsiveness of the splenic T cells to IL-2. These LLC-LN7-bearing mice were also examined for whether cells that phenotypically resemble GM-progenitor cells (ER-MP12+ cells) infiltrate the tumor mass. ER-MP12+ cells composed approximately 10% of the cells isolated from dissociated tumors of mice that had been treated with placebo or with either IFN or TNF alone, but IFN/TNF therapy markedly reduced the number of tumor-infiltrating ER-MP12+ suppressor cells. The IFN/TNF treatment to eliminate GM-suppressor cells and restore T cell responsiveness to IL-2 was next coupled with low dose IL-2 therapy (100 U twice daily). Addition of IL-2 to the treatment regimen did not significantly influence the effectiveness of the IFN/TNF treatment in eliminating GM-suppressor cells from the LLC-LN7 tumor mass. However, inclusion of IL-2 with the IFN/TNF treatment regimen enhanced the CD8+, but not the CD4+, cell content within the tumor, and diminished the number of metastatic lung nodules within the mice. When these tumors were excised, dissociated, and bulk-cultured with a low dose of IL-2, an increased level of cytotoxic T lymphocyte (CTL) activity was generated in the TIL cultures from mice that had received IFN/TNF plus IL-2 treatments. A lesser but detectable level of CTL activity was generated in TIL cultures from mice that were treated with only IFN/TNF, while no CTL activity was generated in tumor cultures from mice receiving only placebo or low-dose IL-2. These results suggest the effectiveness of IFN plus TNF therapy in restoring IL-2 responsiveness in mice bearing GM-suppressor cell-inducing tumors and at enhancing both the intratumoral CD8+ cell content and the generation of CTL activity in bulk cultures of these tumors.This study was supported by the Medical Research Service of the Department of Veterans Affairs, by grants CA-45080 and CA-48080 from the National Institutes of Health, and by the American Cancer Society, Illinois 相似文献
4.
Ichiro Tayasu Toshio Nakamura Hirotaka Oda Fujio Hyodo Yoko Takematsu Takuya Abe 《Ecological Research》2002,17(2):195-206
Stable (13C and 15N) and radio- (14C, 137Cs and 210Pb) isotopes were determined for termites that
have been sampled from a dry evergreen forest in Thailand. A wood-feeding termite, Microcerotermes crassus, was separated from soil-feeders: Termes propinquus, Termes comis and Dicuspiditermes makhamensis by 13C and 15N values. The Termes group in Thailand had less diverse values in 13C and 15N than those in Australia, where the feeding habits of the Termes group are more diverse. Other soil-feeding termites produced similar 13C values, but a larger range in 15N values. 14C-percent modern carbon (pMC) values suggest that the soil-feeding termites used younger carbon than the wood-feeding termites, and this was consistent with the termites from Cameroon, central Africa. Values of 13C and 14C-pMC indicate that surface soil was used by a soil-feeding termite, D. makhamensis, in making the nest mounds, and deeper soil (10–30 cm) by a fungus-growing termite, Macrotermes carbonarius. 210Pb and 137Cs were scarcely incorporated into the termites, although 214Pb was recovered from the workers. The results suggest that stable- and radioisotopes are useful in the study of detritivorous animals, organic matter decomposition and ecosystem engineering.Takuya Abe - deceased. 相似文献
5.
Pre-incubation of somatic tissues of the cut rose Carefree Beauty and miniature roses Red Sunblaze and Baby Katie in 10, 100, or 200 M 2,4-D induced the development of highly rhizogenic callus. Upon transfer of rhizogenic callus to a regeneration medium containing 23 M TDZ and 3 M GA3, a low frequency of shoot organogenesis (3.3%) and somatic embryogenesis (6.6%) was observed. Incubation of leaf and stem internodes in 11, 27, 54, 81, or 108 M NAA followed by transfer of explants to the regeneration medium resulted in up to a 25% increase in shoot organogenesis from callus-derived internodal explants of Red Sunblaze, but no somatic embryogenesis was observed. The influence of glucose versus sucrose in the regeneration medium on leaf explants of Carefree Beauty and Baby Katie pre-incubated in 100 M 2,4-D revealed a difference in genotypic response to shoot organogenesis and somatic embryogenesis. For Carefree Beauty, a concentration of 111 mM glucose induced higher frequencies of both organogenic (33%) and embryogenic calluses (25%) than either 59 mM or 117 mM sucrose. For Baby Katie, no significant difference was found for number of organogenic calluses induced on 59 mM sucrose and 111 mM glucose.Abbreviations ABA
Abscisic acid
- BA
Benzyladenine
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- GA3
Gibberellic acid
- IAA
Indoleacetic acid
- MS
Murashige & Skoog (1962)
- NAA
-Naphthaleneacetic acid
- TDZ
Thidiazuron 相似文献
6.
TOSHIHIRO MIYAJIMA YOSHIHIRO YAMADA EITARO WADA TAKUO NAKAJIMA TADATOSHI KOITABASHI YUKO T. HANBA KOICHI YOSHII 《Biogeochemistry》1997,36(2):205-221
The vertical distribution of dissolved greenhouse gases (CH4,CO2 and N2O), NO
,and 13C of CO2 in Lake Biwa during a stagnantperiod was precisely determined. CO2 as well as NO
was accumulated in the hypolimnion, whereas NO
and CH4concen\-trations were generally higher in theepilimnion than in the hypolimnion. In August, NO
andCH4 were ephemerally accumulated at the thermocline. Theconcentration of CH4 always exceeded equilibrium with respectto air/water exchange. N2O was rather uniformly distributed inboth time and space, and remained near equilibrium with respect toair/water exchange. All of these observations are similar to otherstratified, oligotrophic lakes, in which the hypolimnia were welloxygenated. The 13C of CO2 became morenegative with increasing depth, and showed a strong negativecorrelation with apparent oxygen utilization. From the data, the 13Cvalue of organic matter decomposed into CO2 inthe hypolimnion was calculated by isotope mass-balance, and found tobe in a similar range to 13C of phytoplankton and benthic algaeand distinctively higher than 13C of both terrestrial andsedimentary organic matters. This suggests that autochthonous organicmatter was the major source of CO2. 相似文献
7.
Summary The and subunit of RNA polymerase are thought to be controlled by a translational feedback mechanism regulated by the concentration of RNA polymerase holoenzyme. To study this regulation in vivo, an inducible RNA polymerase overproduction system was developed. This system utilizes plasmids from two incompatibility groups that carry RNA polymerase subunit genes under lac promoter/operator control. When the structural genes encoding the components of core RNA polymerase (, and ) or holoenzyme (, , and 70) are present on the plasmids, induction of the lac promoter results in a two fold increase in the concentration of functional RNA polymerase. The induction of RNA polymerase overproduction is characterized by an initial large burst of synthesis followed by a gradual decrease as the concentration of RNA polymerase increases. Overproduction of RNA polymerase in a strain carrying an electrophoretic mobility mutation in the rpoB gene results in the specific repression of synthesis off the chromosome. These results indicate that RNA polymerase feedback regulation controls synthesis in vivo. 相似文献
8.
The kinetics and temperature dependencies of development and relaxation of light-induced absorbance changes caused by deepoxidation of violaxanthin to antheraxanthin and zeaxanthin (Z; peak at 506 nm) and by light scattering (S; peak around 540 nm) as well as of nonphotochemical quenching of chlorophyll fluorescence (NPQ) were followed in cotton leaves. Measurements were made in the absence and the presence of dithiothreitol (DTT), an inhibitor of violaxanthin deepoxidase. The amount of NPQ was calculated from the Stern-Volmer equation. A procedure was developed to correct gross AS (Sg) for absorbance changes around 540 nm that are due to a spectral overlap with Z. This protocol isolated the component which is caused by light-scattering changes alone (Sn). In control leaves, the kinetics and temperature dependence of the initial rate of rise in Sn that takes place upon illumination, closely matched that of Z. Application of DTT to leaves, containing little zeaxanthin or antheraxanthin, strongly inhibited both Sn and NPQ, but DTT had no inhibitory effect in leaves in which these xanthophylls had already been preformed, showing that the effect of DTT on An and NPQ results solely from the inhibition of violaxanthin deepoxidation. The rates and maximum extents of Sn and NPQ therefore depend on the amount of zeaxanthin (and/or antheraxanthin) present in the leaf. In contrast to the situation during induction, relaxation of Z upon darkening was much slower than the relaxation of Sn and NPQ. The relaxation of Sn and NPQ showed quantitatively similar kinetics and temperature dependencies (Q10=2.4). These results are consistent with the following hypotheses: The increase in lumen-proton concentration resulting from thylakoid membrane energization causes deepoxidation of violaxanthin to antheraxanthin and zeaxanthin. The presence of these xanthophylls is not sufficient to cause Sn or NPQ but, together with an increased lumen-proton concentration, these xanthophylls cause a conformational change, reflected by Sn. The conformational change facilititates nonradiative energy dissipation, thereby causing NPQ. Membrane energization is prerequisite to conformational changes in the thylakoid membrane and resultant nonradiative energy dissipation but the capacity for such changes in intact leaves is quite limited unless zeaxanthin (and/or antheraxanthin) is present in the membrane. The sustained Sn and NPQ levels that remain after darkening may be attributable to a sustained high lumen-proton concentration.Abbreviations A
antheraxanthin
- DTT
dithiothreitol
- F, Fm
chlorophyll fluorescence yield at actual, full closure of the PSII centers
- NPQ
nonphotochemical chlorophyll fluorescence quenching
- PFD
photon flux density
- PSII
photosystem II
- V
violaxanthin
- Z
zeaxanthin
- Sn, Z
spectral absorbance change caused by light-scattering, violaxanthin deepoxidation
We thank Connie Shih for skillful assistance in growing the plants, and for conducting HPLC analyses. A Carnegie Institution Fellowship and a Feodor-Lynen-Fellowship by the Alexander von Humboldt-Foundation to W. B. is gratefully acknowledged. This work was supported in part by Grant No. 89-37-280-4902 of the Competitive Grants Program of the U.S. Department of Agriculture to O.B. This is C. I. W. — D. P. B. Publication No. 1094. 相似文献
9.
ArelA+ strain ofE. coli with four amino acid requirements was starved separately for each amino acid, after which the levels of polysomes, guanosine-5-diphosphate-3-diphosphate and the residual net synthesis of RNA were determined. The polysome level and guanosine-5-diphosphate-3-diphosphate production were coordinately affected by starvation for the different amino acids, whereas no correlation was found between these two parameters and residual RNA synthesis. The main conclusion stemming from these results is that guanosine-5-diphosphate-3-diphosphate cannot act as the sole effector molecule in stringent control of RNA synthesis. 相似文献
10.
Summary The relative rates of RNA polymerase biosynthesis in Bacillus subtilis has been examined under steady-state growth conditions. The synthesis of RNA polymerase subunits (, , , ) has been followed by subunit fractionation of immunoprecipitated [3H]-labelled samples on SDS-polyacrylamide gels. The stoichiometries of ::: subunits have been determined from cultures pulse-labelled during steady-state growth. The results suggest that an unassembled pool of the -subunit exists from which the holoenzyme is formed.Upon shift-up from acetate to glycerol containing medium, a rapid rise in the differential rate of core enzyme synthesis was observed, while the rate of synthesis of the -subunit was not stimulated. During shift-down, a concomitant reduction in the rate of synthesis of all subunits occurred for the first 20 min after the shift; thereafter, a rate of synthesis characteristic of the new growth rate was established.As cultures enter sporulation, an immediate reduction in the rate of -subunit synthesis was demonstrated. 相似文献
11.
Water and nitrogen dynamics in an arid woodland 总被引:5,自引:0,他引:5
Arid environments are characterized by spatial and temporal variation in water and nitrogen availability. differences in 15N and D of four co-occurring species reveal contrasting patterns of plant resource acquisition in response to this variation. Mineralization potential and nitrogen concentration of surface soils associated with plant canopies were greater than inter-canopy locations, and values decreased with increasing depth in both locations. Mineralization potential and nitrogen concentration were both negatively correlated with soil 15N. The spatial variation in soil 15N caused corresponding changes in plant 15N such that plant 15N values were negatively correlated with nitrogen concentration of surface soils. Plants occurring on soils with relatively high nitrogen concentrations had lower 15N, and higher leaf nitrogen concentrations, than plants occurring on soils with relatively low nitrogen concentrations. Two general temporal patterns of water and nitrogen use were apparent. Three species (Juniperus, Pinus andArtemisia) relied on the episodic availability of water and nitrogen at the soil surface. 15N values did not vary through the year, while xylem pressure potentials and stem-water D values fluctuated with changes in soil moisture at the soil surface. In contrast,Chrysothamnus switched to a more stable water and nitrogen source during drought. 15N values ofChrysothamnus increased throughout the year, while xylem pressure potentials and stem-water D values remained constant. The contrasting patterns of resource acquisition have important implications for community stability following disturbance. Disturbance can cause a decrease in nitrogen concentration at the soil surface, and so plants that rely on surface water and nitrogen may be more susceptible than those that switch to more stable water and nitrogen sources at depth during drougnt. 相似文献
12.
P. A. Dibrov R. L. Lazarova V. P. Skulachev M. L. Verkhovskaya 《Journal of bioenergetics and biomembranes》1989,21(3):347-357
Addition of Na+ to the K+-loadedVibrio alginolyticus cells, creating a 250-fold Na+ gradient, is shown to induce a transient increase in the intracellular ATP concentration, which is abolished by the Na+/H+ antiporter, monensin. The pNa-supported ATP synthesis requires an additional driving force supplied by endogenous respiration or, alternatively, by a K+ gradient (high [K+] inside). In the former case, ATP formation is resistant to the protonophorous uncoupler. Dicyclohexylcarbodiimide and diethylstilbestrol, but not vanadate, completely inhibit Na+ pulse-induced ATP formation. The data agree with the assumption that Na+-ATP-synthase is involved in oxidative phosphorylation inV. alginolyticus. Interrelation of H+ and Na+ cycles in bacteria is discussed.Abbreviations
and
electrochemical gradients of H+ and Na+, respectively
-
transmembrane electric potential difference
- pH, pNa, and pK
concentration gradients of H+, Na+, and K+, respectively
- CCCP
carbonyl cyanidem-chlorophenylhydrazone
- DCCD
N,N-dicyclohexylcarbodiimide
- DES
diesthylstilbestrol
- HQNO
2-heptyl-4-hydroxyquinolineN-oxide
- Tricine
N[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine 相似文献
13.
I. A. Bass Zh. M. Gorlenko O. N. Danilevskaya A. D. Dmitriev E. S. Kalyaeva S. Z. Mindlin A. V. Timopheev R. S. Khesin 《Molecular & general genetics : MGG》1977,154(1):101-110
Summary In merodiploid cells containing a double dose of structural genes of RNA polymerase subunits-rpoBand rpoC-the rate of and subunits synthesis is 2 times higher than in haploidcells. Missence mutation rpoC1 (tsX) alters polypeptide and inducesthe and subunits synthesis at increased rate, particularly at a nonpermissive temperature. When rpoBCoperon carrying mutation rpoC1 is duplicated no dosage effect is observed. In the rpoC
+/rpoC1 heterodiploid the rpoC1 mutation does not significantly accelerate RNA polymerase subunits synthesis i.e. is recessive with respect to rpoC
+
Rifampicin causes 6-fold stimulation of RNA polymerase subunits synthesis in a sensitive wild-type strain. The rpoC1 mutation itself accelerates the synthesis of these subunits 3-fold. In the presence of rifampicin the mutant strain produces 13–22-fold faster as compared to wild-type strain without the drug. Thus, the effects of rifampicin and the mutation are multiplied suggesting that these factors act independently. Similar data have also been obtained with rifampicin-treated cells of rpoB22 (ts22) amber-mutant.After UV-irradiation of cells and synthesis is depressed much stronger than the total protein synthesis. Infection with a transducing phage rif
d-47 which carries rpoB gene provokes a higher rate of synthesis. When pre-irradiated cells (500 erg/mm2) are infected with this phage, the rate of synthesis grows 20-fold compared to irradiated, non-infected cells and 6.5-fold compared to intact cells.The data are discussed in terms of the possible regulatory mechanisms of RNA polymerase subunit synthesis. 相似文献
14.
S. H. Chiou P. Azari M. E. Himmel H. K. Lin W. P. Chang 《Journal of Protein Chemistry》1989,8(1):19-32
A detailed investigation of the hydrodynamic and aggregation behaviors has been made on the -crystallins of bovine lens. Results from this study indicated that H (high-molecular-weight -crystallin) and L (low-molecular-weight -crystallin) exhibited considerable heterogeneity in their native structures and subunit polypeptides. Low-speed sedimentation equilibrium showed a heterogeneous paucidisperse system in each -crystallin fraction. Viscosity and circular dichroism studies pointed to a compact and globular shape and the presence of -sheet and -turns in these crystallins. Dissociation of H by urea and guanidinium HCl followed by reassociation during gel-filtration chromatography produced an elution pattern with two fractions corresponding to L crystallin and high-molecular-weight aggregates without the formation of native H. By contrast, under similar treatment, about 60% L reassociated into the correct native structure and the rest into high-molecular-weight fractions. Amino acid analyses of H and L and their corresponding subunit polypeptides demonstrated the close similarity of these crystallins. Trace element analyses indicated that both Ca and Mg are present in H and L crystallins and may be involved in maintaining the native quarternary structures of these proteins. 相似文献
15.
Raymond Delpech V Ihara M Coddou C Matsuda K Sattelle DB 《Invertebrate neuroscience : IN》2003,5(1):29-35
Nereistoxin (NTX), a natural neurotoxin from the salivary glands of the marine annelid worm Lumbriconereis heteropoda, is highly toxic to insects. Its synthetic analogue, Cartap, was the first commercial insecticide based on a natural product. We have used voltage-clamp electrophysiology to compare the actions of NTX on recombinant nicotinic acetylcholine receptors (nicotinic AChRs) expressed in Xenopus laevis oocytes following nuclear injection of cDNAs. The recombinant nicotinic AChRs investigated were chicken 7, chicken 42 and the Drosophila melanogaster/chicken hybrid receptors SAD/2 and ALS/2. No agonist action of NTX (0.1–100 M) was observed on chicken 7, chicken 42 and the Drosophila/chicken hybrid nicotinic AChRs. Currents elicited by ACh were reduced in amplitude by NTX in a dose-dependent manner. The toxin was slightly more potent on recombinant Drosophila/vertebrate hybrid receptors than on vertebrate homomeric (7) or heteromeric (42) nicotinic AChRs. Block by NTX of the chicken 7, chicken 42 and the SAD/2 and ALS/2 Drosophila/chicken hybrid receptors is in all cases non-competitive. Thus, the site of action on nicotinic AChRs of NTX, to which the insecticide Cartap is metabolised in insects, differs from that of the major nicotinic AChR-active insecticide, imidacloprid. 相似文献
16.
We examined the effects of fertilizer application, especially the effects of fertigation and types of fertilizer (inorganic and organic) on yields and 15N and 13C values of tomato (Lycopersicon esculentum Mill. cv. Saturn). Fertigation is a method in which an appropriate diluted liquid fertilizer is applied to the plants each time they are drip-irrigated. We developed a method of organic fertigation using corn steep liquor (CSL) as the liquid fertilizer, because it is an industrial byproduct of cornstarch manufacture and can be used very effectively. We compared fruit yield, mineral content, 15N value, and 13C value of tomatoes grown under three different fertilizer treatments, basal dressing: basal dressing with granular chemical fertilizer; inorganic fertigation: fertigation with liquid chemical fertilizer; and organic fertigation: fertigaion with CSL. Mineral contents of tomatoes grown with basal dressing were generally lower than those grown under either fertigation treatment. These results indicated that yields and mineral contents were influenced more by the method of fertilizer application than by whether the fertilizers were inorganic or organic. There were, however, significant differences in the 15N values of tomato fruits grown under different types of fertilizer applications, especially between inorganic and organic fertilizers. The 15N value of the chemical fertilizer used for basal dressing was 0.81 ± 0.45{}, that of the chemical fertilizer for fertigation was 0.00 ± 0.04{}, and that of CSL was 8.50 ± 0.71{}. The 15N values of the soils reflected the 15N values of the fertilizers. Moreover, the 15N values of the fruits corresponded to the 15N values of the applied fertilizers. The 15N values were 3.18 ± 1.34{} in the fruits grown with a basal dressing of chemical fertilizer, 0.30 ± 0.61 in those grown under inorganic fertigation, and 7.09 ± 0.68 in those grown under organic fertigation. On the other hand, although the 13C values in the soil also reflected the 13C values of the applied fertilizers, there was no significant difference in the 13C values of fruits among the different treatments. In conclusion, because the 15N values of fertilizers correlated well with those of the fruits, it may be possible to use 15N values as an indicator of organic products. 相似文献
17.
Summary Insulin binding to human placenta membranes treated at pH 7.6 or 8.5 in the presence or absence of 2.0mm DTT for 5 min, followed by the simultaneous removal of the DTT and pH adjustment to pH 7.6, displayed curvilinear (heterogeneous) insulin binding plots when analyzed by the method of Scatchard. However, Triton X-100 solubilization followed by Bio-Gel A-1.5m gel filtration chromatography of the placenta membranes previously treated with DTT at pH 8.5 generated a nearly straight line (homogeneous) Scatchard plot.125I-insulin affinity crosslinking studies coupled with Bio-Gel A-1.5m gel filtration chromatography demonstrated that the alkaline pH and DTT treatment of placenta membranes followed by detergent solubilization generated an heterodimeric insulin receptor complex from the 22 heterotetrameric disulfide-linked state. The ability of alkaline pH and DTT to produce a functional heterodimeric insulin receptor complex was found to be time dependent with maximal formation and preservation of tracer insulin binding occurring at 5 min. These data demonstrate that (i) a combination of alkaline pH and DTT treatment of placenta membranes can result in the formation of a functional heterodimeric insulin receptor complex. (ii) the heterodimeric complex displays homogeneous insulin binding. (iii) the insulin receptor membrane environment maintains the 22 association state, which displays heterogeneous insulin binding, despite reduction of the critical domains that are responsible for the covalent interaction between the heterodimers.Abbreviations used are ATP
adenosine 5-triphosphate
- DTT
dithiothreitol
- SDS
sodium dodecyl sulfate
- DSS
disuccinimidyl suberate
- NEM
N-ethylmaleimide
- IGF-I
insulin-like growth factor-I
- EDTA
ethylenediaminetetraacetic acid
- HEPES
4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid 相似文献
18.
Multiple-quantum 2D and 3D bi-directional HCNCH experiments are presented for the correlation of base and ribose protons/carbons in 13C/15N labeled HIV-1 TAR RNA. In both 2D and 3D experiments, the magnetization of H1 is transferred to H6/H8 and H1 through H1-C1-N1/9-C6/8-H6/8 and H1-C1-N1/9-C1-H1 pathways, and the magnetization of H6/8 is transferred to H1 and H6/8 through H6/8-C6/8-N1/9-C1-H1 and H6/8-C6/8-N1/9-C6/8-H6/8 pathways. Chemical shifts of four different nuclei (H1, C1, C6/8 and H6/8) are sampled in the 2D experiment. The correlation of base and ribose protons/carbons is established by the rectangular arrangement of crossover and out-and-back peaks in the proton/carbon correlated spectrum. The rectangular connections can be further resolved using the nitrogen dimension in a 1H/13C/15N 3D experiment. Furthermore, by taking advantage of the well separated chemical shifts of N1 (pyrimidine) and N9 (purine), the 2D spectrum can be simplified into two sub-spectra based on their base type. Both experiments were tested on a 13C/15N labeled 27-mer HIV-1 TAR RNA containing a UUCG hairpin loop. 相似文献
19.
Frank Sundler M.D. R. E. Carraway R. Håkanson J. Alumets M. P. Dubois 《Cell and tissue research》1978,194(3):367-376
Summary Two distinct populations of endocrine cells in the chicken thymus display neurotensin and somatostatin immunoreactivity, respectively. Both cell types are few in number at hatching but proliferate rapidly during the first week. The neurotensin cells are Grimelius-positive and Hellerström-Hellmannegative. The somatostatin cells are Grimelius-negative and Hellerström-Hellman-positive. Both cell populations are non-argentaffin. The somatostatin-like material extracted from chicken thymus behaves immunochemically and chromatographically similar to synthetovine somatostatin, while the neurotensin-like material, from the thymus as well as from the gut, differs from synthetic bovine neurotensin in that it appears larger in size and more basic. 相似文献
20.
N Nakada K Yoshinaga A Ishihama H Nagasawa-Fujimori 《Molecular & general genetics : MGG》1982,188(2):173-178
Summary On exposure to high temperature of a temperature-sensitive RNA polymerase subunit (rpoC92) mutant of Escherichia coli, selective reduction was observed in the rate of synthesis of a group of proteins including RNA polymerase subunit. The finding that the synthesis of subunit but not subunit was specifically repressed in this mutant grown at non-permissive temperature indicates that the functionally intact RNA polymerase is required for the synthesis of subunits be coordinated. In addition, the assembly of newly synthesized RNA polymerase subunits was inefficient in this mutant at the steps where altered subunit was involved, and the unassembled enzyme subunits were rapidly and preferentially degraded. During recovery to non-restricted growth, the synthesis of both and subunits was transiently enhanced in parallel leading to recovery of the intracellular concentration of functional RNA polymerase. 相似文献