期刊
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
卷 1830, 期 8, 页码 4189-4201出版社
ELSEVIER
DOI: 10.1016/j.bbagen.2013.03.027
关键词
Sanguinarine; Human telomeric DNA; NHE III1 upstream of the P1 promoter of c-myc; Fluorescence and CD spectroscopy; Calorimetry; Telomerase repeat amplification protocol
资金
- Molecular Mechanism of Diseases and Drug Action, Department of Atomic Energy, Govt. of India
- CSIR, India [HCP 004]
Background: Interaction of putative anticancer agent sanguinarine with two quadruplex forming sequences, human telomeric DNA (H24) and NHE III1 upstream of the P1 promoter of c-myc (Pu27), has been studied to understand the structural basis of the recognition. Methods: Absorption, fluorescence and circular dichroism spectroscopy have been employed to characterize the association. Energetics of the interaction was studied by isothermal titration and differential scanning calorimetry. TRAP assay was done to assess the inhibitory potential of sanguinarine. Results: Absorption and fluorescence studies show that sanguinarine has high binding affinity of similar to 10(5) M-1 for both sequences. Binding stoichiometry is 2:1 for H24 and 3:1 for Pu27. Results suggest stacking interaction between planar sanguinarine moiety and G-quartets. Circular dichroism spectra show that sanguinarine does not cause structural perturbation in the all-parallel Pu27 but causes a structural transition from mixed hybrid to basket form at higher sanguinarine concentration in case of H24. The interaction is characterized by total enthalpy-entropy compensation and high heat capacity values. Differential scanning calorimetry studies suggest that sanguinarine binding increases the melting temperature and also the total enthalpy of transition of both quadruplexes. TRAP results show that sanguinarine effectively blocks telomerase activity in a concentration dependent manner in cell extracts from MDAMB-231 breast cancer cell lines. Conclusion: These results suggest that there is a difference in the structural modes of association of sanguinarine to the quadruplexes. General significance: It helps to understand the role of quadruplex structures as a target of small molecule inhibitors of telomerase. (c) 2013 Elsevier B.V. All rights reserved.
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