Induction of a Remarkable Conformational Change in a Human Telomeric Sequence by the Binding of Naphthyridine Dimer: Inhibition of the Elongation of a Telomeric Repeat by Telomerase
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- 作者：Kazuhiko Nakatani ; Shinya Hagihara ; Shinsuke Sando ; Shigeru Sakamoto ; Kentaro Yamaguchi ; Chihaya Maesawa ; and Isao Saito
- 刊名：Journal of the American Chemical Society
- 出版年：2003
- 出版时间：January 22, 2003
- 年：2003
- 卷：125
- 期：3
- 页码：662 - 666
- 全文大小：192K
- 年卷期：v.125,no.3(January 22, 2003)
- ISSN：1520-5126
文摘
The binding of a dimeric form of the 2-amino-1,8-naphthyridine derivative (naphthyridine dimer)to a human telomeric sequence, TTAGGG, was investigated by UV melting, CD spectra, and CSI-MSmeasurements. Both the 9-mer d(TTAGGGTTA) and the 15-mer d(TTAGGGTTAGGGTTA) showedapparent melting temperatures (Tm) of 45.6 and 63.6 
C, respectively, in the presence of naphthyridinedimer (30 
M) in sodium cacodylate buffer (50 mM, pH 7.0) containing 100 mM NaCl. The CD spectra at235 and 255 nm of the 9-mer increased in intensity accompanied with strong induced CDs at 285 and 340nm upon complex formation with naphthyridine dimer. UV titration of the binding of naphthyridine dimer tothe 9-mer at 320 nm showed a hypochromism of the spectra. A Scatchard plot of the data showed thepresence of multiple binding sites with different association constants. Cold spray ionization massspectrometry of the complex between naphthyridine dimer and the 9-mer clearly showed that one to threemolecules of the ligand bound to the dimer duplex of the 9-mer. Telomeric repeat elongation assay showedthat the binding of naphthyridine dimer to the telomeric sequence inhibits the elongation of the sequenceby telomerase.
C, respectively, in the presence of naphthyridinedimer (30 M) in sodium cacodylate buffer (50 mM, pH 7.0) containing 100 mM NaCl. The CD spectra at235 and 255 nm of the 9-mer increased in intensity accompanied with strong induced CDs at 285 and 340nm upon complex formation with naphthyridine dimer. UV titration of the binding of naphthyridine dimer tothe 9-mer at 320 nm showed a hypochromism of the spectra. A Scatchard plot of the data showed thepresence of multiple binding sites with different association constants. Cold spray ionization massspectrometry of the complex between naphthyridine dimer and the 9-mer clearly showed that one to threemolecules of the ligand bound to the dimer duplex of the 9-mer. Telomeric repeat elongation assay showedthat the binding of naphthyridine dimer to the telomeric sequence inhibits the elongation of the sequenceby telomerase.