@article {https://doi.org/10.1002/elps.202100077, title = {Quantitative characterization of human oncogene promoter G-quadruplex DNA-ligand interactions using a combination of mass spectrometry and capillary electrophoresis}, journal = {ELECTROPHORESIS}, volume = {42}, number = {n/a}, year = {2021}, month = {05/2021}, pages = {1450-1460}, type = {Research}, chapter = {1450}, abstract = {
Abstract Human c-KIT oncogene is known to regulate cell growth and proliferation, and thus, acts as a probable target in the treatment of gastrointestinal tumors (GIST). To identify small molecule ligands which can specifically bind with the G-quadruplex (G4) in the c-KIT promoter region as potential antitumor agents, we propose the combination of electrospray ionization\–mass spectrometry (ESI-MS), capillary electrophoresis frontal analysis (CE-FA), and Taylor dispersion analysis (TDA) to accurately investigate the G4/ligands binding properties. First, ESI\–MS was used for initial screening of natural products (NPs). CE-FA was then used to calculate specific binding constants and the stoichiometry of the native state binding pair in solution. Next, TDA, a micro-capillary flow technique was used to examine the effect of the ligand binding on the diffusivity and particle size of the c-KIT G4. Two of the screened NPs, scopolamine butylbromide (L1) and isorhamnetin-3-O-neohesperidoside (L3), were found to specifically bind to the c-KIT G4 with binding constants of around 104 M\–1 and 1:1 stoichiometry in a free solution. TDA data showed that ligand binding (both L1 and L3) induced the c-KIT strands to fold into a tightly structured G4 with a decreased hydrodynamic radius. These ligands have the potential to be drug candidates for the regulation of c-KIT gene transcription by stabilizing the G4 structure. This methodology not only increased the speed of analysis but also improved its accuracy and specificity compared with the conventional binding approaches.
}, keywords = {binding constant, c-KIT oncogene, Capillary electrophoresis frontal analysis, Electrospray ionization{\textendash}mass spectrometry, G-quadruplex, Taylor dispersion analysis}, doi = {https://doi.org/10.1002/elps.202100077}, url = {https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/abs/10.1002/elps.202100077}, author = {Wang, Shuangshuang and Yang, Yang and Yang, Yunhe and Li, Huihui and Chen, David D.Y.} } @article {doi:10.1002/elps.201800049, title = {Pressure-assisted capillary electrophoresis frontal analysis for faster binding constant determination}, journal = {ELECTROPHORESIS}, volume = {39}, number = {14}, year = {2018}, month = {07/2018}, pages = {1786-1793}, abstract = {Abstract Adding external pressure during the process of capillary electrophoresis usually add to the band broadening, especially if the pressure induced flow is significant. The resolution is normally negatively affected in pressure-assisted capillary electrophoresis (PACE). Frontal analysis (FA), however, can potentially benefit from using an external pressure while avoiding the drawbacks in other modes of CE. In this work, possible impact from the external pressure was simulated by COMSOL Multiphysics\®. Under a typical CE-FA set-up, it was found that the detected concentrations of analyte will not be significantly affected by an external pressure less than 5\ psi. Besides, the measured ligand concentration in PACE-FA was also not affected by common variables (molecular diffusion coefficient (10\−8 to 10\−11 m2/s), capillary length etc). To provide an experimental proof, PACE-FA is used to study the binding interactions between hydroxypropyl β-cyclodextrin (HP-β-CD) and small ligand molecules. Taking the HP-β-CD /benzoate pair as an example, the binding constants determined by CE-FA (18.3\ \±\ 0.8 M\−1) and PACE-FA (16.5\ \±\ 0.5\ M\−1) are found to be similar. Based on the experimental results, it is concluded that PACE-FA can reduce the time of binding analysis while maintaining the accuracy of the measurements.
}, keywords = {Capillary electrophoresis frontal analysis, Cyclodextrin complex, DNA quadruplex, Pressure assisted CEFA}, doi = {10.1002/elps.201800049}, url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/elps.201800049}, author = {Qian Cheng and Wang Su and Fu Hengqing and Turner Robin F. B. and Li Huihui and Chen David D. Y.} } @article {ELPS:ELPS6128, title = {Mobility-based correction for accurate determination of binding constants by capillary electrophoresis-frontal analysis}, journal = {ELECTROPHORESIS}, volume = {38}, year = {2017}, pages = {1572-1581}, type = {Research}, keywords = {Capillary electrophoresis frontal analysis, Improved binding constant accuracy, Mobility corrected equation}, issn = {1522-2683}, doi = {10.1002/elps.201600450}, url = {http://dx.doi.org/10.1002/elps.201600450}, author = {Qian, Cheng and Kovalchik, Kevin A. and MacLennan, Matthew S. and Huang, Xiaohua and Chen, David D.Y.} }