Purification of Carotenoids and Monoterpen Aldehydes from Iranian Saffron and Investigation of Their Effect on the Structure of DNA, Histone H1 and H1-DNA Complex

Bathaie, S.Z.; Ashrafi, M.; Bolhasani, A.; Etemadi-Kia, B.; Moosavi-Movahedi, A.A.

Document Type : Research Paper

Authors

¹ Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

² Institiute of Biochemistry & Biophysics, University of Tehran, Tehran, Iran.

Abstract

Saffron (Crocus Sativus L.) has different biological properties including anticancer activity. In the present research, we studied the possible molecular mechanisms of the saffron as an anticancer compound (interaction with DNA, H1 and H1-DNA complex), in addition to the purification of saffron components. The isolation and purification of major components of Iranian Saffron including carotenoids (crocin, crocetin and dimethylcrocetin) and monoterpen aldehydes (picrocrocin and safranal) performed by different methods. The existence of three types of picrocrocin and four types of crocin in Iranian saffron by using TLC and HPLC techniques was shown. The major fraction of crocins and picrocrocins were extracted. The effects of saffron components on two macromolecules, DNA and H1, as well as the H1-DNA complex were studied. The interaction of saffron extract and all of the cited components investigated with DNA. The results showed the non-intercalative binding of the mentioned ligands with DNA and suggested the minor groove binding. The binding parameters for crocin interaction with DNA obtained (n= 0.54 and K= 2 × 10 ^-5 M^-1). The circular dichroism spectra indicated that the mentioned components interact with DNA at low concentrations, which is accompanied by some conformational changes, B- to C-DNA transition; however, at higher concentrations, they induce more conformational changes, i.e. unstacking of DNA bases. Spectrophotometric investigations of histone H1 titration by different components showed some changes in the peaks. These data indicated the binding of these components to histone H1. All components quenched the fluorescence emission of histone H1 as the result of its structural changes upon complex formation. By Scatchard analysis of the quenching data the binding parameters (n and K) calculated. Histone H1- DNA interaction were also studied in the absence and presence of each ligand. Our results showed that both carotenoids and monoterpene aldehydes of saffron cause the reduction on H1-DNA interaction by about 5 to 22%. These results are preliminary data to confirm our hypothesis that may be the anticancer effect of saffron is due to their interaction with DNA and reduction in the H1-DNA complxes.

Keywords

References

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Iranian Journal of Medicinal and Aromatic Plants Research

Purification of Carotenoids and Monoterpen Aldehydes from Iranian Saffron and Investigation of Their Effect on the Structure of DNA, Histone H1 and H1-DNA Complex

References

References

Volume 22, Issue 2 - Serial Number 32
September 2006
Pages 85-97

Purification of Carotenoids and Monoterpen Aldehydes from Iranian Saffron and Investigation of Their Effect on the Structure of DNA, Histone H1 and H1-DNA Complex

References

References

Volume 22, Issue 2 - Serial Number 32September 2006Pages 85-97

Volume 22, Issue 2 - Serial Number 32
September 2006
Pages 85-97