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A homogeneous phase protein-based assay for the high throughput screening of drugs was developed using enhanced green fluorescent protein (EGFP) as the reporter. For that, a fusion protein between calmodulin (CaM) and EGFP was constructed in order to monitor the conformational changes induced in CaM upon binding to tricyclic anti-depressant drugs. In the presence of Ca2+, CaM undergoes a conformational change exposing a hydrophobic pocket that interacts with CaM-binding proteins, peptides, and drugs. Further, the conformational changes induced in CaM upon binding to Ca2+ and the target analyte drug, leads to a change in the microenvironment of EGFP concomitant with a change in its fluorescence intensity. The observed change in fluorescence intensity of EGFP can be correlated to the concentration of the analyte present in the sample. Further, the response of CaM–EGFP fusion protein in the presence of Ca2+ to increasing concentrations of phenothiazines and structurally related tricyclic anti-depressants was investigated. Dose-response curves for various tricyclic anti-depressants were prepared. Moreover, this assay can serve as a model system for other homogeneous binding assays for pharmaceuticals employing genetically fused binding proteins with reporter proteins and may find applications in the high throughput screening of tricyclic anti-depressants. 相似文献
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Jurva U Bissel P Isin EM Igarashi K Kuttab S Castagnoli N 《Journal of the American Chemical Society》2005,127(35):12368-12377
Single-electron transfer and hydrogen atom transfer pathways have been proposed to account for the cytochrome P450-catalyzed alpha-carbon oxidations of amines. With the aid of electrochemistry-electrospray ionization mass spectrometry, the electrochemical potentials required for the one-electron oxidations of N-methyl- and selected N-cyclopropyl-4-phenyl-1,2,3,6-tetrahydropyridinyl derivatives and the chemical fates of the resulting aminyl radical cations have been investigated. Comparison of the results of these studies with those observed in the corresponding enzyme catalyzed oxidations suggests that aminyl radical cations are not obligatory intermediates in the cytochrome P450-catalyzed alpha-carbon oxidations of this class of substrates. 相似文献
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In this paper, using power series methods we give an abstract Korovkin type approximation theorem for a sequence of positive linear operators mapping \({C\left(X, \mathbb{R}\right)}\) into itself. 相似文献
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Various aniline derivatives were synthesized by selective reduction of aryl azides in the presence of a dichloro(p-cymene)ruthenium(II) dimer ([Ru(p-cymene)Cl2]2) via hydrolysis of sodium borohydride. The hydrogenation reactions were carried out in aqueous media at room temperature. Most of the reactions were completed within 10 min with quantitative yields. 相似文献
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Journal of Global Optimization - Channel coding aims to minimize the errors that occur during the transmission of digital information from one place to another. Low-density parity-check codes can... 相似文献