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Carp growth hormone (cGH) cDNA, in which Cys-123 was mutated to Ala, was prepared, transferred to the expression vector, expressed in Escherichia coli and the mutant was purified to homogeneity. The mutation only slightly improved yield of the monomeric fraction, indicating that Cys-123 is not involved in improper refolding. As compared to cGH, the mutant (cGH-C123A) exhibited lower binding affinity toward homologous liver receptors and lower bioactivity in a 3T3-F442A preadipocyte bioassay despite the fact that both hormones exhibited almost identical cross-reactivity with anti-cGH antibodies. These results, along with those of a structural comparison to hGH, suggest that Cys-123 is located in the hydrophobic core of the hormone, and is most likely affecting the conformation of the binding site. Dimeric forms of the hormone and its mutant were less active than their respective monomers. Homologous binding experiments using a carp liver microsomal fraction revealed a single receptor population with Kd = 0.77 nM and Bmax = 241 fmol/mg microsomal protein.
Résumé Un ADN complémentaire (cDNA) de l'hormone de croissance de carpe (cGH), dans lequel l'acide aminé Cys-123 a été muté en Ala, a été préparé, inséré dans un vecteur d'expression, et exprimé dans Escherichia coli. Le mutant a ensuite été purifié jusqu'à homogénéité. La mutation améliore seulement faiblement la production de la fraction monomérique, indiquant que le Cys-123 n'est pas impliqué dans un repliement erroné. Comparé à la cGH, sa forme mutée (cGH-C123A) montre une plus faible affinité de liaison vis à vis de récepteurs hépatiques homologues, et une plus faible activité biologique dans un test réalisé sur des préadipocytes 3T3-F442A; cela en dépit du fait que les deux hormones présentent des réactions croisées presque identiques avec un anticorps anti-cGH polyclonale. Ces résultats, associés à une comparaison à la structure de l'hGH, suggèrent que le Cys-123 est localisé dans la partie hydrophobique de l'hormone, et affecte, le plus vraisemblablement, la conformation du site de liaison. Les formes dimériques de l'hormone et de sa forme mutée sont moins actives que leurs monomères respectifs. Les études de liaison homologue, réalisées avec des fractions microsomales de foie, révèlent une population unique de récepteurs de Kd = 0,77 nM et de Bmax = 241 fmol/mg de proteine microsomale.
  相似文献   
3.
Arieh Levy 《Euphytica》1985,34(3):811-815
Summary Papaver bracteatum is a potential crop for the codeine industry. Seed shattering prevailing in the species is a major problem which reduces the seed yields. A spontaneous mutant with capsules having no pores of dehiscence was isolated in an experimental field. The seed yields of this shattering-resistant mutant are lower than in plants with normal capsules but the thebaine content of the capsules from the mutant is higher. The mutation is monogenic dominant; it can be used for breeding cultivars for dual purpose-thebaine and seeds.  相似文献   
4.
The ubiquitination of the hypoxia-inducible factor (HIF) by the von Hippel-Lindau tumor suppressor (pVHL) plays a central role in the cellular response to changes in oxygen availability. pVHL binds to HIF only when a conserved proline in HIF is hydroxylated, a modification that is oxygen-dependent. The 1.85 angstrom structure of a 20-residue HIF-1alpha peptide-pVHL-ElonginB-ElonginC complex shows that HIF-1alpha binds to pVHL in an extended beta strand-like conformation. The hydroxyproline inserts into a gap in the pVHL hydrophobic core, at a site that is a hotspot for tumorigenic mutations, with its 4-hydroxyl group recognized by buried serine and histidine residues. Although the beta sheet-like interactions contribute to the stability of the complex, the hydroxyproline contacts are central to the strict specificity characteristic of signaling.  相似文献   
5.
Cyhexatin, dienochlor and four experimental acaricides, U-25322 (1-benzoyl-3-phenylhydrazine), U-27415 1-benzoyl-3-(2, 4, 6-trichlorophenyl)hydrazine, U-42558 [N1(4-chloro-o-tolyl) N2-methyl-N1-(phenylthio)formamidine], and U-42564 [N2-methyl-N1-(phenylthio)- N1-(2, 4-xylyl)formamidine], at 125 mg litre-1, induced spindown by the carmine spider mite, Tetranychus cinnabarinus (Boisduval). Cyhexatin caused the largest number of mites to spin-down, dienochlor the least. Spinning occurred only during the first 6 h after treatment. Mites spun the same average length of threads in response to all compounds. The pattern of spin-down was even and similar for all the compounds except dienochlor for which it was sporadic. Acaricide-induced spin-down is postulated to be an ‘all-or-nothing’ reaction.  相似文献   
6.
The present study was undertaken to examine the effect of recombinant chicken leptin administered to fed ad libitum and feed-restricted immature chickens of a layer strain on ovarian development and the timing of sexual maturity. In the first experiment 11-week-old pullets (77 days of age) fed ad libitum were injected daily with leptin at four dose levels (4, 16, 64 and 256 microg/kg body weight) until sexual maturity (lay of the first egg). Leptin treatment at the highest dose significantly (P<0.05) advanced the onset of puberty (day 116.3+/-1.0) in comparison to controls (day 121.3+/-1.2). The rises of luteinizing hormone, estradiol and progesterone in blood plasma were also advanced by leptin treatment. In the second experiment, both full-fed and feed-restricted pullets (79 days of age) were injected daily with leptin (256 microg/kg body weight). In birds fed ad libitum, exogenous leptin again significantly (P<0.05) advanced first ovipostion (day 118.4+/-1.4 versus day 124.4+/-1.7), while abolishing the significant (P<0.05) delay caused by feed restriction (day 131.5+/-1.6) and restoring the normal onset of sexual maturity (day 125.7+/-1.6). Analysis of the ovaries in 106-day-old pullets revealed that leptin injections advanced follicular development, particularly in birds fed ad libitum, and significantly (P<0.01) reduced follicular apoptosis both in full-fed and feed-restricted birds. In conclusion, we have shown that in female chickens exogenous leptin advances the onset of puberty by attenuation of ovarian apoptosis and enhancement of folliculogenesis.  相似文献   
7.
Chicken leptin: properties and actions   总被引:6,自引:0,他引:6  
Chicken leptin cDNA shows a high homology to mammalian homologous, with an expression localized in the liver and adipose tissue. It is noteworthy, that the hepatic expression is most likely associated with the primary role that this organ plays in lipogenic activity in avian species. As in mammals, chicken leptin expression is regulated by hormonal and nutritional status. This regulation is tissue-specific and with a high sensitivity in the liver compared to adipose tissue. The blood leptin levels are regulated by the nutritional state with high levels in the fed state compared to the fasted state. The recombinant chicken leptin markedly inhibits food intake as reported in mammals, suggesting the presence of an hypothalamic leptin receptor. The chicken leptin receptor has been identified and all functional motifs are highly conserved compared to mammalian homologous. Chicken leptin receptor is expressed in the hypothalamus but also in other tissues such as pancreas, where leptin inhibits insulin secretion and thus may have a key role in regulating nutrient utilization in this species.  相似文献   
8.
A chicken leptin-specific radioimmunoassay   总被引:3,自引:0,他引:3  
Recombinant chicken leptin was used to produce an antiserum in order to develop a specific and sensitive radioimmunoassay (RIA) for chicken leptin in plasma and serum. We have used either murine or chicken leptin as tracer and competition curves were performed using recombinant chicken leptin. Variations in leptin plasma levels in different chicken strains and various nutritional states were correlated with the physiological status. Leptin plasma concentrations were regulated by the nutritional state with higher levels in the fed state as compared to the fasted state (3.36 +/- 0. 13 versus 2.78 +/- 0.11 ng/ml) and being dependent upon the age. Higher leptin levels were found in 22 week-old as compared to 15 week-old layer chickens (2.709 +/- 0.172 versus 1.478 +/- 0.102 ng/ml). We have also shown that the multispecies leptin RIA kit (LINCO Inc.) underestimated leptinemia compared to the chicken leptin- specific RIA reported here. In conclusion the RIA developed in the present study is specific to the chicken and thus may be considered as powerful tool for investigating the physiological significance of leptin in chickens.  相似文献   
9.
Pomegranate peel is a nutritive-rich byproduct whose amounts are extensively growing due to the exponential increase in the production of pomegranate juice and "ready to eat" arils. Pomegranate peel is a rich source for antioxidants and thus may serve in the prevention of cattle diseases and in the improvement of beef products, making it an attractive component in beef cattle diets. The present study aims to evaluate the effect of commonly used storage practices on the nutritive and antioxidative properties of pomegranate peel. In general, storage conditions preserved most antioxidant capacity. Ensiling ambivalently affected the nutritive values of the peel and promoted increased levels of antioxidative components. In addition to polyphenols, nonphenolic components, such as alpha- and gamma-tocopherols, contributed to the total antioxidative capacity, and several minerals found in the peel added to its nutritional value. Dietary supplementation with fresh peels promoted significant increases in feed intake and alpha-tocopherol concentration in the plasma, with positive tendency toward increased weight gain of bull calves. All in all, the nutritive value and the antioxidant capacity of pomegranate peel turn it into a favorable health-promoting constituent of feedlot beef cattle diet.  相似文献   
10.
Migration of different mineral particles within columns of soil‐sand mixtures containing 10 or 20 mass % of soil was investigated by establishing differences in the mineral suite between the ”︁bulk clay” and the ”︁mobile fine material” fractions. The ”︁bulk clay” fractions of all soils contained smectite, palygorskite, kaolinite, quartz, feldspar, and calcite. The soils were saturated with sodium by leaching with NaCl solution, and then leached with distilled water. Clay dispersion and particle migration occurred in the columns. Values of SAR (sodium adsorption ratio) of the effluent decreased with time due to carbonate dissolution. At a certain SAR value, the clays apparently formed aggregates, and as a consequence particle migration stopped in the column. In addition to clay‐sized particles (< 2 μm), very‐fine‐silt‐sized particles (2— 5 μm) were able to migrate in the soil‐sand mixtures, too, and to some extent fine‐silt‐sized particles (5—10 μm) as well. Average size of mobile particles decreases with increase of soil content in the soil‐sand mixtures. The mineralogical composition of the ”︁mobile fine material” changed during the experiment. At the beginning of the experiment, the ”︁mobile fine material” was enriched in the non‐phyllosilicates (especially in calcite, and in some cases in quartz, feldspar and dolomite) and contained low concentrations of phyllosilicates (smectite, palygorskite and kaolinite). At the end of the experiment, the proportion of non‐phyllosilicates decreased, and as a consequence, the proportion of phyllosilicates increased. Among the non‐phyllosilicates, calcite was the most mobile mineral. Among the phyllosilicates, palygorskite was preferentially mobilized in topsoil horizons. In subsoil horizons, on the other hand, kaolinite was preferentially mobilized. This difference was explained by the different nature of carbonates in the topsoil and subsoil horizons. Palygorskite is preferentially occluded within the soil carbonates of lacustrine origin over smectite and kaolinite. These carbonates are present mainly in the subsoil horizons. As a consequence, the presence of these carbonates in the subsoil horizons decreases the migration of mainly palygorskite.  相似文献   
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