PMID- 23771525 OWN - NLM STAT- MEDLINE DCOM- 20131021 LR - 20191210 IS - 1618-2650 (Electronic) IS - 1618-2642 (Linking) VI - 405 IP - 19 DP - 2013 Jul TI - Targeting human c-Myc promoter duplex DNA with actinomycin D by use of multi-way analysis of quantum-dot-mediated fluorescence resonance energy transfer. PG - 6271-80 LID - 10.1007/s00216-013-7041-8 [doi] AB - Actinomycin D (Act D), an oncogenic c-Myc promoter binder, interferes with the action of RNA polymerase. There is great demand for high-throughput technology able to monitor the activity of DNA-binding drugs. To this end, binding of 7-aminoactinomycin D (7AAD) to the duplex c-Myc promoter was investigated by use of 2D-photoluminescence emission (2D-PLE), and the resulting data were subjected to analysis by use of convenient and powerful multi-way approaches. Fluorescence measurements were performed by use of the quantum dot (QD)-conjugated c-Myc promoter. Intercalation of 7AAD within duplex base pairs resulted in efficient energy transfer from drug to QD via fluorescence resonance energy transfer (FRET). Multi-way analysis of the three-way data array obtained from titration experiments was performed by use of restricted Tucker3 and hard trilinear decomposition (HTD). These techniques enable analysis of high-dimensional and complex data from nanobiological systems which include several spectrally overlapped structures. It was almost impossible to obtain robust and meaningful information about the FRET process for such high overlap data by use of classical analysis. The soft approach had the important advantage over univariate classical methods of enabling us to investigate the source of variance in the fluorescence signal of the DNA-drug complex. It was established that hard trilinear decomposition analysis of FRET-measured data overcomes the problem of rank deficiency, enabling calculation of concentration profiles and pure spectra for all species, including non-fluorophores. The hard modeling approach was also used for determination of equilibrium constants for the hybridization and intercalation equilibria, using nonlinear fit data analysis. The intercalation constant 3.6 x 10(6) mol(-1) L and hybridization stability 1.0 x 10(8) mol(-1) L obtained were in good agreement with values reported in the literature. The analytical concentration of the QD-labeled DNA was determined by use of nonlinear fitting, without using external standard calibration samples. This study was a successful application of multi-way chemometric methods to investigation of nano-biotechnological systems where several overlapped species coexist in solution. FAU - Gholami, Somayeh AU - Gholami S AD - Department of Chemistry, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan, Iran. FAU - Kompany-Zareh, Mohsen AU - Kompany-Zareh M LA - eng PT - Evaluation Study PT - Journal Article DEP - 20130616 PL - Germany TA - Anal Bioanal Chem JT - Analytical and bioanalytical chemistry JID - 101134327 RN - 0 (Fluorescent Dyes) RN - 0 (MYC protein, human) RN - 0 (Proto-Oncogene Proteins c-myc) RN - 1CC1JFE158 (Dactinomycin) RN - 7240-37-1 (7-aminoactinomycin D) RN - 9007-49-2 (DNA) SB - IM MH - DNA/*chemistry/genetics/metabolism MH - Dactinomycin/*analogs & derivatives/*chemistry/metabolism MH - Fluorescence Resonance Energy Transfer/instrumentation/*methods MH - Fluorescent Dyes/chemistry MH - Humans MH - *Promoter Regions, Genetic MH - Proto-Oncogene Proteins c-myc/*chemistry/genetics/metabolism EDAT- 2013/06/19 06:00 MHDA- 2013/10/22 06:00 CRDT- 2013/06/18 06:00 PHST- 2013/02/24 00:00 [received] PHST- 2013/05/02 00:00 [accepted] PHST- 2013/04/22 00:00 [revised] PHST- 2013/06/18 06:00 [entrez] PHST- 2013/06/19 06:00 [pubmed] PHST- 2013/10/22 06:00 [medline] AID - 10.1007/s00216-013-7041-8 [doi] PST - ppublish SO - Anal Bioanal Chem. 2013 Jul;405(19):6271-80. doi: 10.1007/s00216-013-7041-8. Epub 2013 Jun 16.