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Mechanism of activation of the ret proto-oncogene by multiple endocrine neoplasia 2A mutations. 总被引:19,自引:0,他引:19
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Transforming activity of the c-ret proto-oncogene with multiple endocrine neoplasia (MEN) 2A mutations was investigated by transfection of NIH 3T3 cells. Mutant c-ret genes driven by the simian virus 40 or cytomegalovirus promoter induced transformation with high efficiencies. The 170-kDa Ret protein present on the cell surface of transformed cells was highly phosphorylated on tyrosine and formed disulfide-linked homodimers. This result indicated that MEN 2A mutations induced ligand-independent dimerization of the c-Ret protein on the cell surface, leading to activation of its intrinsic tyrosine kinase. In addition to the MEN 2A mutations, we further introduced a mutation (lysine for asparaginic acid at codon 300 [D300K]) in a putative Ca(2+)-binding site of the cadherin-like domain. When c-ret cDNA with both MEN 2A and D300K mutations was transfected into NIH 3T3 cells, transforming activity drastically decreased. Western blot (immunoblot) analysis revealed that very little of the 170-kDa Ret protein with the D300K mutation was expressed in transfectants while expression of the 150-kDa Ret protein retained in the endoplasmic reticulum was not affected. This result also demonstrated that transport of the Ret protein to the plasma membrane is required for its transforming activity. 相似文献
24.
Yamazaki Y Hosono K Matsuda H Minami N Asai M Nakanishi H 《Biotechnology and bioengineering》1991,38(10):1218-1222
Twelve species of optically active metallocene derivatives having a 4-nitrophenyl group were prepared with horse liver alcohol dehydrogenase- or lipase-catalyzed optical resolution as the key step. The second harmonic generation (SHG) efficiently of these products was measured by the power method using the fundamental light from the Nd:YAG laser. (-)-1- (4-Nitrophenylthio)ethylferrocene (9), (+)-1-(4-nitrophenylthio)ethylruthenocene (16), (+)-1-(4-nitrophenylthio)ethylosmocene (19), (+) -1-(5-nitro-2-pyridylthio)ethylruthenocene (21), and (+) -1-[(4-nitrophenylhydrazono)methyl] -2-methylferrocene (12) showed SHG signals. The highest SHG efficiency was found with (+) -16, being 27 times more intense than the commonly used urea standard. 相似文献
25.
Kiyofumi Asai Takayoshi Hirano Shuji Kaneko Akihiko Moriyama Keiko Nakanishi Ichiro Isobe Yaman Z. Eksioglu Taiji Kato 《Journal of neurochemistry》1992,59(1):307-317
Neurofibroma tissue was investigated for the presence of glial growth modulators that would suppress the proliferation of glial cells. A novel endogenous polypeptide inhibitor of proliferation and DNA synthesis in glial cells, gliostatin, was purified from the extracts of neurofibroma by a procedure comprising dye and anion-exchange column chromatography, and HPLC. A monoclonal antibody raised against partially purified gliostatin showed no cross-reactivity with known cytokines, but adsorbed the growth inhibitory activity of gliostatin and immunochemically visualized the putative gliostatin bands on western blot analyses. Although the product showed an apparent M(r) of 100,000 accompanied by an inhibitory activity on gel filtration column chromatography, it migrated at a lower apparent M(r) of 50,000 under the reducing conditions on western blotting, indicating that a homodimeric structure of native gliostatin consisted of 50-kDa subcomponents. Gliostatin was a potent growth inhibitor acting at nanomolar concentrations against all glial tumor cells and glia maturation factor-stimulated astroblasts, but not neuronal cells. 相似文献
26.
The translational diffusion coefficient (D) of H-meromyosin in actin (F-actin) and ATP solution was measured under conditions wherein the actin-activated ATPase activity is close to its maximal value at a very low electrolyte concentration. The results were compared with similar data obtained with 0.1 M KCl, where H-meromyosin and actin were almost completely dissociated. With 0.1 M KCl, it was found that there was no dependence of the D of H-meromyosin on actin concentration. On the other hand, at a very low electrolyte concentration, it was found that the D of H-meromyosin did depend on actin concentration; at a rather high actin concentration (and activation of ATPase), it was slightly larger than at low or zero actin concentrations. This behavior of D at a low electrolyte concentration is interpreted on the assumption that even in solution, H-meromyosin molecules can actively slide on actin filaments due to the ATPase activity. 相似文献
27.
A tentative and phenomenological analysis of negative electric birefringence, which has often been observed as an anomalous birefringence phenomenon in a concentrated solution of rodlike macromolecules, is presented. Tobacco mosaic virus (TMV) was used as a typical example for the investigation. It was found that if the applied electric field is sufficiently high, the steady-state birefringence becomes becomes positive even at a very high concentration of TMV. From this finding and analysis of the time course of birefringence transients, it was suggested that the TMV (common strain, OM type), which originally has no inherent permanent dipole, behaves as if it possesses a permanent dipole perpendicular to its long axis. Supporting evidence was also obtained from birefringence experiments on concentrated solutions of the HR strain of TMV, which has an inherent permanent dipole along its long axis. Other possibilities, for example, the effects of the walls of electrodes or of polymerization of TMV molecules, were excluded. 相似文献
28.
Female rats were irradiated with 190R of X-rays at 10 days of age and sacrificed 4, 7 or 12 months later. Their ovaries were histologically examined and serum levels and pituitary contents of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were determined by radioimmunoassay. Both serum levels and pituitary contents of LH and FSH rose significantly 4 and 7 months after irradiation, although the ovaries were markedly reduced in weight. On the contrary, 12 months after irradiation, the ovaries increased in weight and consisted mostly of polyhedral, hyperplastic interstitial cell masses, and both LH and FSH in the serum and pituitary were reduced to normal levels. These characteristic changes in the ovarian weight and histological appearance could not be observed in the similarly irradiated animals which were received daily injections of estrone for the first 30 days of postnatal life, i.e., daily injections of 50 mug for the first 10 days, 100 mug for the middle 10 days and 200 mug for the last 10 days. Serum LH levels of the estrogenized irradiated rats at 7 or 12 months of age did not elevate although those of FSH were significantly higher than the non-irradiated intact levels. From these results, a rise in the blood levels of LH and the FSH may be attributed to the increase in weight and the histological changes in the ovaries of the irradiated female rats, and the elevation of only FSh level may not result in the abnormal growth of the irradiated ovaries. 相似文献
29.
The fluorescence properties of chlorophyll a and b monomolecular films at the air-water interface were measured by a high sensitivity fluorophotometer using the photon-counting method. The fluorescence intensity of chlorophyll molecules in monomolecular films in the absence of any diluents did not decrease simply with the mean distance of chlorphyll molecules. Over the range of the mean distances from 27 to 21 A, three fluorescence components (peaks at 685, 695 and 715 nm) of chlorophyll a were observed. In the case of chlorophyll beta, two fluorescence components (peaks at 667 and 685 nm) were observed over the range of the mean distances from 34 to 24 A. When the mean distance was 18 A, the short wavelength component of chlorophyll beta disappeared, and only the long avelength component was observed. 相似文献
30.
Effects of sodium, potassium and chloride ions on the membrane potential of Valonia aegagropila 总被引:1,自引:0,他引:1
Changes of the resting potential of Valonia cell in sea wateragainst a 10-fold increase of the external concentrations ofK$, Na$ and Cl were 1±1, 6.2±0.1 and 38.9±4mV, respectively. The potassium conductance was smaller than7 µ/cm2, while the Na and Cl conductances were 45 and281 µ/cm2, respectively, in normal sea water. The positivevacuolar potential could be explained by these ionic conductances.On the other hand, the membrane became more sensitive to K$,if the cell was incubated for about 30 min in K-rich (100 mM)sea water. It is worth noting, however, that the membrane conductancewas lower in the K-rich sea water than in the normal sea water. (Received October 7, 1974; ) 相似文献