Structural analysis of telomere G-quartet and molecular design of new nucleic acid aptamer

端粒G四联体结构分析及新型核酸适体分子设计

• 批准号: 15570098
• 负责人: KANAORI Kenji
• 金额: $ 2.3万
• 依托单位: Kyoto Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570098/
• 关键词: telomere; four stranded structure; nucleic acid aptamer; i-moitf; G-quartet; NMR

The P-chiral stereodefined phosphorothioate groups have been introduced into all the four internucleotide positions of d(T_<PS1>C_<PS2>C_<PS3>C_<PS4>C) (PSn : phosphorothioate group), and among possible 16 diastereomers of PS-d(TC_4), 10 stereomers have been synthesized to investigate the effects of the sense of the P-chirality upon the structure and stability of the i-motif structure. The temperature dependence of circular dichroism spectra showed melting temperature (Tm) of the [all R_p]PS-d(TC_4) i-motif was 31 ℃ identical to that of the parent oligomer, PO-d(TC_4), while that of the [all S_p]PS-d(TC_4) i-motif was largely decreased by 11℃. Single substitution of R_p with S_p caused a decrease of T_m by 3〜4℃ at positions of PS1, 2, and 3, and by 1℃ at that of PS4, showing the additive property of the T_m suppression. The comparison of the NOESY spectra between [all R_p]PS-, [all S_p]PS-, and PO-d(TC_4) showed that intra-residual H6-H3' and H2"-H4' NOE cross-peaks of the [all S_p] isomer are weaker than those of the [all R_p] isomer and PO-d(TC_4), indicating the change in the C3'-endo conformation and glycosidic bond angle. The structural alternation for the i-motif formed by [all S_p]PS-d(TC_4) is also suggested by the chemical shift differences of C2/C3/C4 H2"and H4' protons from those of [all R_p]PS-d(TC_4) and PO-d(TC_4). These results suggest that the S_p configuration at phosphorus of the phosphorothioate linkage change the sugar-phosphorothioate conformation and intermolecular interaction in the narrow groove, leading to the destabilization of the i-motif structure.

P-手性立体定向的硫代磷酸酯基团已被引入d（T_<PS1>C_<PS2>C_<PS3>C_<PS4>C）的所有四个核苷酸间位置（PSn：在PS-d（TC_4）的16种可能的非对映异构体中，合成了10种立体异构体，以研究P-手性的意义对i-基序结构和稳定性的影响。圆二色谱的温度依赖性表明，[all R_p]PS-d（TC_4）i-motif的解链温度（Tm）与母体寡聚体PO-d（TC_4）相同，为31 ℃，而[all S_p]PS-d（TC_4）i-motif的解链温度则降低了11℃。S_p单取代R_p，使PS_1、2、3位的Tm降低3 ~ 4℃，PS_4位的Tm降低1℃，表现出Tm抑制的加和性。[all R_p]PS-、[all S_p]PS-和PO-d（TC_4）的NOESY谱比较表明，[all S_p]异构体的H_6-H_3 '和H_2”-H_4' NOE交叉峰比[all R_p]异构体和PO-d（TC_4）的弱，表明C_3 '-内构象和糖苷键角发生了变化。[all S_p]PS-d（TC_4）和PO-d（TC_4）的C_2/C_3/C_4 H_2 ~+和H_4 ~+质子的化学位移差异也表明了[all S_p] PS-d（TC_4）形成的i基序的结构变化。这些结果表明，硫代磷酸酯键磷位的S_p构型改变了糖-硫代磷酸酯构象和窄沟中的分子间相互作用，导致i-基序结构的不稳定。

项目成果

Semihemoglobins, high oxygen affinity dimeric forms of human hemoglobin respond efficiently to allosteric effectors without forming tetramers.

半血红蛋白是人血红蛋白的高氧亲和力二聚体形式，可有效响应变构效应物，而不形成四聚体。

• DOI: 10.1074/jbc.m405909200
• 发表时间: 2004
• 期刊: The Journal of biological chemistry
• 作者: Tsuneshige,Antonio;Kanaori,Kenji;Samuni,Uri;Danstker,David;Friedman,JoelM;Neya,Saburo;Giangiacomo,Laura;Yonetani,Takashi
• 通讯作者: Yonetani,Takashi

Two-Metal Ion, Ni(II) and Cu(II), Binding a-Helical Coiled-coil Peptide.

双金属离子，Ni(II) 和 Cu(II)，结合 a-螺旋卷曲螺旋肽。

• 发表时间: 2004
• 期刊: Journal of American Chemical Society 126
• 作者: Tanaka;T.;Mizuno;T.;Fukui;S.;Hiroaki;H.;Oku;J.;Kanaori;K.;Tajima;K. Shirakawa;M.
• 通讯作者: M.

Yamada, M.: "Nitration of 2-deoxyguanosine by a NO/O2 gas mixture : identification and"Org.Lett.. 5. 3173-3176 (2003)

Yamada, M.：“NO/O2 气体混合物对 2-脱氧鸟苷的硝化：鉴定和”Org.Lett.. 5. 3173-3176 (2003)

Solution structure of marinostatin, a natural ester-linked protein protease inhibitor

• DOI: 10.1021/bi048034x
• 发表时间: 2005-02-22
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Kanaori, K;Kamei, K;Hara, S
• 通讯作者: Hara, S

Effect of phosphorothioate chirality on i-motif structure and stability.

• DOI: 10.1021/bi035419r
• 发表时间: 2004-05
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: K. Kanaori;S. Sakamoto;H. Yoshida;P. Guga;W. Stec;K. Tajima;K. Makino