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1.
Solid‐state 31P NMR investigation on the status of guanine nucleotides in paclitaxel‐stabilized microtubules 下载免费PDF全文
Ga Hyang Lee Su Young Oh Kwon Joo Yeo Taslima Ferdous Meeyeon Cho Younkee Paik 《Magnetic resonance in chemistry : MRC》2015,53(5):330-336
Microtubule dynamics is a target for many chemotherapeutic drugs. In order to understand the biochemical effects of paclitaxel on the GTPase activity of tubulin, the status of guanine nucleotides in microtubules was investigated by 31P cross‐polarization magic angle spinning (CPMAS) NMR. Microtubules were freshly prepared in vitro in the presence of paclitaxel and then lyophilized in sucrose buffer for solid‐state NMR experiments. A 31P CPMAS NMR spectrum with the SNR of 25 was successfully acquired from the lyophilized microtubule sample. The broadness of the 31P spectral lines in the spectrum indicates that the molecular environments around the guanine nucleotides inside tubulin may not be as crystalline as reported by many diffraction studies. Deconvolution of the spectrum into four spectral components was carried out in comparison with the 31P NMR spectra obtained from five control samples. The spectral analysis suggested that about 13% of the nucleotides were present as GTP and 37% as GDP in the β‐tubulin (E‐site) of the microtubules. It was found that most of the GDPs were present as GDP‐Pi complex in the microtubules, which seems to be one of the effects of paclitaxel binding. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
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We investigated the biomaterial and pharmaceutical utility of pure silk fibroin (SF) protein as a possible for separation, using Sephadex G-25 gel filtration chromatography and simply preparing SF microsphere particles (SFMP) by spray dryer. Also, some of its physicochemical properties and morphology were investigated. Obtaining microspheres and/or submicronic particles by spray dryer method was accelerated or completed with the transition from the random coil to the β-sheet structure during spray dryer treatment. It was identified by the basic Fourier transform infrared spectroscopy of SFMP. The various pH range of SFMP’s swelling ratio is dependent on the pH of the solution, not on the occurred gelation. Morphologically, SFMP was spherical in shape, and particles, average 2±10 μm in size, were observed by scanning electron microscope and particle analyzer, respectively. The average molecular weight (MW) of pure SF protein dissolved in calcium chloride is about 61,500 g/mol as measured by gel permeation chromatography. 相似文献
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Jeong JO Byun J Jeon ES Gwon HC Lim YS Park J Yeo SJ Lee YJ Kim S Kim DK 《Experimental & molecular medicine》2002,34(4):278-284
Direct injection of the vascular endothelial growth factor (VEGF) gene plasmid DNA into the myocardium was shown to induce development of new blood vessels to increase the circulation in the heart of patients with coronary artery diseases. However, such angiogenic gene therapy (via naked DNA) was limited by low level of gene expression. Furthermore, the temporal and spatial characteristics of VEGF gene transfer in the heart are not known. In this study, we demonstrated that a plasmid vector, containing the human cytomegalovirus immediate early (HCMV IE) promoter and enhancer, induces greater expression of gene in the rat heart monitored by gene fused to the chloramphenicol acetyl transferase (CAT) reporter, than four different viral and cellular promoters. Interestingly, expression of VEGF121 protein showed an earlier peak, a shorter duration, and a wider distribution than that of CAT only. Therefore, a plasmid vector with an HCMV IE promoter/enhancer provides clear advantages over other previously developed plasmids. Furthermore, expression profile of VEGF121 gene may provide useful information in the design of angiogenic gene therapy in the heart. 相似文献
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An organopalladium complex containing ortho-metalated (S)-(1-(dimethylamino)ethyl)naphthalene as the chiral auxiliary has been used to promote the asymmetric hydrophosphination reactions between diphenylphosphine and (E)- or (Z)-diphenyl-1-propenylphosphine in high regio- and stereoselectivities under mild conditions. Hydrophosphination of (Z)-diphenyl-1-propenylphosphine with diphenylphosphine gave (S)-(-)-prophos as the major product. Using the same chiral metal template, the corresponding hydrophosphination reaction with (E)-diphenyl-1-propenylphosphine gave (R)-(+)-prophos predominantly. The hydrophosphination reactions generated the asymmetric diphosphines as bidentate chelates on the chiral naphthylamine palladium templates. The template products obtained undergo cis-trans isomerization in solution to form an equilibrium mixture of regioisomers. X-ray analysis of the major template products obtained from the hydrophosphination of (Z)-diphenyl-1-propenylphosphine reveals that the two regioisomers are cocrystallized in a 1:1 ratio. The naphthylamine auxiliary could be removed chemoselectively from the template products by treatment with concentrated hydrochloric acid to form the corresponding optically pure neutral complexes [(R)- or (S)-(prophos)PdCl(2)]. Subsequently, the (R)- and (S)-dichloro complexes undergo ligand displacement with aqueous cyanide to generate the corresponding optically pure diphosphine ligands in high yields. Mechanistic pathways explaining the stereoselectivity of the chiral organopalladium template promoted hydrophosphination reactions are also proposed. 相似文献
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Both adaptive and deleterious responses of cells to ethanol are likely triggered by short-term interactions of the cells with ethanol. Many studies have demonstrated the direct effect of ethanol on growth factor-stimulated cell proliferation. Using Swiss 3T3 cells whose growth was inhibited by ethanol in a concentration-dependent manner, we further investigated the molecular mechanisms of acute ethanol treatment by examining its effect on EGF- and PDGF-mediated cellular signaling systems for the mitogenic function. Tyrosine autophosphorylation of the growth factor receptors was partially prevented by ethanol in intact cells. When ethanol was included before or after EGF stimulation, no effect on the receptor signaling was observed. Here we also report that ethanol inhibits activation of ERK induced by both EGF and PDGF. EGF-induced JNK activation was reduced but PDGF-induced rapid JNK activation was delayed by the addition of ethanol. The balance between its inhibitory and stimulatory effect on the signaling molecules might determine the rate of cell growth. 相似文献
8.
R. Westwood Dudley H. Williams Adrian N. H. Yeo 《Journal of mass spectrometry : JMS》1970,3(12):1485-1488
The [NO2C7H6]+ ions generated from m-alkylnitrobenzenes have been shown to be different in their decomposition from those generated from p-alkylnitrobenzenes, even when the alkyl group is methyl and the departing fragment a hydrogen radical. Thus, in these cases even molecular ions of relatively high internal energy do not reversibly ring-expand to cycloheptatriene structures. In addition, the [NO2C7H6]+ ions, assumed to be benzylic, do not ring-expand to nitrotropylium ions at internal energies sufficient to cause subsequent loss of NO or NO2 from the p- and m-isomers, respectively. 相似文献
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