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ABSTRACT: A three-dimensional pharmacophore model was generated utilizing a set of known inhibitors of c-Myc-Max heterodimer formation. The model successfully identified a set of structurally diverse compounds with potential inhibitory activity against c-Myc. Nine compounds were tested in vitro, and four displayed affinities in the micromolar range and growth inhibitory activity against c-Myc-overexpressing cells. These studies demonstrate the applicability of pharmacophore modeling to the identification of novel and potentially more puissant inhibitors of the c-Myc oncoprotein.
Journal of Medicinal Chemistry 03/2009; 52(5):1247-50. · 4.80 Impact Factor
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ABSTRACT: The basic-helix-loop-helix-leucine-zipper domains of the c-Myc oncoprotein and its obligate partner Max are intrinsically disordered (ID) monomers that undergo coupled folding and binding upon heterodimerization. We have identified the binding sites and determined the structural means by which two unrelated small molecules, 10058-F4 and 10074-G5, bind c-Myc and stabilize the ID monomer over the highly ordered c-Myc-Max heterodimer. In solution, the molecules bind to distinct regions of c-Myc and thus limit its ability to interact with Max and assume a more rigid and defined conformation. The identification of multiple, specific binding sites on an ID domain suggests that small molecules may provide a general means for manipulating the structure and function of ID proteins, such as c-Myc.
Chemistry & biology 12/2008; 15(11):1149-55. · 6.52 Impact Factor
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ABSTRACT: Compounds that selectively prevent or disrupt the association between the c-Myc oncoprotein and its obligate heterodimeric partner Max (Myc-Max compounds) have been identified previously by high-throughput screening of chemical libraries. Although these agents specifically inhibit the growth of c-Myc-expressing cells, their clinical applicability is limited by their low potency. We describe here several chemical modifications of one of these original compounds, 10058-F4, which result in significant improvements in efficacy. Compared with the parent structure, these analogues show enhanced growth inhibition of c-Myc-expressing cells in a manner that generally correlates with their ability to disrupt c-Myc-Max association and DNA binding. Furthermore, we show by use of a sensitive fluorescence polarization assay that both 10058-F4 and its active analogues bind specifically to monomeric c-Myc. These studies show that improved Myc-Max compounds can be generated by a directed approach involving deliberate modification of an index compound. They further show that the compounds specifically target c-Myc, which exists in a dynamic and relatively unstructured state with only partial and transient alpha-helical content.
Molecular Cancer Therapeutics 10/2007; 6(9):2399-408. · 5.23 Impact Factor
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ABSTRACT: Differences in gene expression between salinity stressed and normally grown wheat seedlings were compared by the differential display (DD) technique. One DD-derived cDNA clone was characterized as a partial sequence of the wheat asparagine ynthetase (AS) gene by sequence analysis and homology search of GenBank databases. Two AS genes of wheat, TaASN1 and TaASN2, were further isolated by the RT-PCR approach. Comparison of the deduced polypeptide of TaASN1 and TaASN2 with AS proteins from other organisms revealed several homologous regions, in particular, the conserved glutamine binding sites and Class-II Glutamine amidotransferases domain. The functionality of TaASN1 was demonstrated by complementing an Escherichia coli asparagine auxotroph. TaASN1 transcripts were detected in roots, shoots, anthers and young spikes by RT-PCR analysis. Abundance of TaASN1 mRNA in young spikes and anthers was higher than that in shoots and roots under normal growth conditions. TaASN1 was dramatically induced by salinity, osmotic stress and exogenous abscisic acid (ABA) in wheat seedlings. TaASN2 transcripts were very low in all detected tissues and conditions and were only slightly induced by ABA in roots.
Journal of Plant Physiology 02/2005; 162(1):81-9. · 2.79 Impact Factor
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ABSTRACT: Pyridoxal kinase (EC 2.7.1.35) is a key enzyme in the conversion of vitamin B6 to pyridoxal 5'-phosphate (PLP). PLP is the crucial cofactor required by numerous enzymes involved in amino acids metabolism. Recently, studies with Arabidopsis salt overly sensitive 4 mutants demonstrated that pyridoxal kinase is a novel salt tolerance determinant important for the regulation of Na+ and K+ homeostasis in plants. We describe here the TaPdxK gene which encodes a pyridoxal kinase, cloned from Triticum aestivum by RACE PCR method. The putative amino acid sequence of TaPdxK is 78% identical to Arabidopsis AtSOS4. Southern analysis suggests that there are at least two copies of pyridoxal kinase genes in wheat genome. The expression of TaPdxK cDNAs complements an Escherichia coli mutant defective in pyridoxal kinase. TaPdxK transcripts were detected in roots, shoots, spikes and anthers by RT-PCR analysis. TaPdxK expression level was not regulated by salt, ABA, and osmotic stress.
Journal of Plant Physiology 10/2004; 161(9):1053-60. · 2.79 Impact Factor
