5'-O-dimethoxytrityl-3'-O-t-butyldimethylsilyl-N²-(phenoxyacetyl)-2′-deoxyguanosine | 642462-83-7

• 分子结构分类: 有机化合物 - 苯类化合物 - 三苯基化合物
• 英文名称: 5'-O-dimethoxytrityl-3'-O-t-butyldimethylsilyl-N²-(phenoxyacetyl)-2′-deoxyguanosine
• 英文别名: 3’-O-(tertbutyldimethylsilyl)-5’-O-(4,4’-dimethoxytrityl)-N2-phenoxyacetyl-2’-deoxyguanosine；3'-O-tert-butyldimethylsilyl-5'-O-dimethoxytrityl-N2-phenoxyacetyl-2'-deoxyguanosine；N-[9-[(2R,4S,5R)-5-[[bis(4-methoxyphenyl)-phenylmethoxy]methyl]-4-[tert-butyl(dimethyl)silyl]oxyoxolan-2-yl]-6-oxo-1H-purin-2-yl]-2-phenoxyacetamide
• CAS: 642462-83-7
• 化学式: C₄₅H₅₁N₅O₈Si
• 分子量: 818.014
• InChiKey: ALQIIMQEVVMUDF-OPWUGURCSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  7.84
• 重原子数：
  59
• 可旋转键数：
  16
• 环数：
  7.0
• sp3杂化的碳原子比例：
  0.33
• 拓扑面积：
  144
• 氢给体数：
  2
• 氢受体数：
  10

反应信息

• 作为反应物：
  描述：

  5'-O-dimethoxytrityl-3'-O-t-butyldimethylsilyl-N²-(phenoxyacetyl)-2′-deoxyguanosine 在 对甲苯磺酸 作用下， 以 甲醇 、 二氯甲烷 为溶剂， 以0.73 g的产率得到N2-phenoxyacetyl-3'-TBDMS-2'-deoxyguanosine
  参考文献：
  名称：

  柔性O6-2'-脱氧鸟苷-亚烷基-O6-2'-脱氧鸟苷内链交联的合成，表征和修复

  摘要：

  作为两个链内交联（IaCL）DNA的模型系统，已经合成了通过两个连续的2'-脱氧鸟苷的O 6原子之间的亚烷基键束缚的寡核苷酸，这些残基之间没有磷酸二酯键。这些丁烯和庚烯连接的寡核苷酸及其互补的DNA序列之间形成的双链体的UV热变性研究表明，相对于未修饰的双链体，其稳定性降低了约20°C。模型IaCL双链体的圆二色性光谱显示了B型DNA的特征，表明病变引起的总体扰动最小。研究了包含IaCL模型的双链体作为O 6的底物来自人和大肠杆菌（Ada-C和OGT）的-烷基鸟嘌呤DNA烷基转移酶（AGT）蛋白。发现人类AGT可以修复两种IaCL双链体，对庚烯和丁烯类似物的修复效率更高，这进一步增加了我们对该蛋白可以修复的底物的了解。

  DOI：

  10.1002/chem.201501103
• 作为产物：
  描述：

  5’-O-dimethoxytrityl-N²-phenoxyacetyl-2’-deoxyguanosine 、 叔丁基二甲基氯硅烷 在 咪唑 、 4-二甲氨基吡啶 作用下， 以 二氯甲烷 为溶剂， 生成 5'-O-dimethoxytrityl-3'-O-t-butyldimethylsilyl-N²-(phenoxyacetyl)-2′-deoxyguanosine
  参考文献：
  名称：

  柔性O6-2'-脱氧鸟苷-亚烷基-O6-2'-脱氧鸟苷内链交联的合成，表征和修复

  摘要：

  作为两个链内交联（IaCL）DNA的模型系统，已经合成了通过两个连续的2'-脱氧鸟苷的O 6原子之间的亚烷基键束缚的寡核苷酸，这些残基之间没有磷酸二酯键。这些丁烯和庚烯连接的寡核苷酸及其互补的DNA序列之间形成的双链体的UV热变性研究表明，相对于未修饰的双链体，其稳定性降低了约20°C。模型IaCL双链体的圆二色性光谱显示了B型DNA的特征，表明病变引起的总体扰动最小。研究了包含IaCL模型的双链体作为O 6的底物来自人和大肠杆菌（Ada-C和OGT）的-烷基鸟嘌呤DNA烷基转移酶（AGT）蛋白。发现人类AGT可以修复两种IaCL双链体，对庚烯和丁烯类似物的修复效率更高，这进一步增加了我们对该蛋白可以修复的底物的了解。

  DOI：

  10.1002/chem.201501103

文献信息

• Site-specific covalent capture of human O⁶-alkylguanine-DNA-alkyltransferase using single-stranded intrastrand cross-linked DNA
  作者：D. K. O'Flaherty、C. J. Wilds

  DOI：10.1039/c6ob02246d

  日期：——

  nsferase (hAGT) to the 3′-end of DNA in excellent yields with short reaction times by using intrastrand cross-linked (IaCL) DNA probes. This strategy exploited the substrate specificity of hAGT to generate the desired DNA–protein covalent complex. IaCL DNA linking two thymidine residues, or linking a thymidine residue to a 2′-deoxyguanosine residue (either in a 5′→3′ or 3′→5′ fashion), lacking a phosphodiester

  据报道，通过使用链内交联（IaCL）DNA探针，可在短时间内以优异的产率将人O 6-烷基鸟嘌呤-DNA-烷基转移酶（hAGT）缀合到DNA的3'-末端。该策略利用了hAGT的底物特异性来生成所需的DNA-蛋白质共价复合物。制备了连接两个胸苷残基或将胸苷残基连接至2'-脱氧鸟苷残基（以5'→3'或3'→5'的方式），在交联位点缺少磷酸二酯键的IaCL DNA使用亚磷酰胺策略，然后进行固相合成。相对于未修饰的对照双链体，所有包含模型IaCL的双链体均显示出热稳定性降低。所述Ô 4 -胸苷-亚烷基- ö评估的任何AGT均无法修复4-胸苷和（5'→3'） O 6 -2'-脱氧鸟苷-亚烷基-O 4-胸苷IaCL DNA加合物（人AGT和大肠杆菌的同源物，OGT和Ada​​- C）。（5'→3'） O 4-胸苷-亚烷基-O 6含有丁烯或庚烯系链的-2'-脱氧鸟苷IaCL DNA已被人类变异体有效修复，
• Synthesis and characterization of an O6-2′-deoxyguanosine-alkyl-O6-2′-deoxyguanosine interstrand cross-link in a 5′-GNC motif and repair by human O6-alkylguanine-DNA alkyltransferase
  作者：Francis P. McManus、Qingming Fang、Jason D. M. Booth、Anne M. Noronha、Anthony E. Pegg、Christopher J. Wilds

  DOI：10.1039/c0ob00093k

  日期：——

  O6-2′-Deoxyguanosine-alkyl-O6-2′-deoxyguanosine interstrand DNA cross-links (ICLs) with a four and seven methylene linkage in a 5′-GNC- motif have been synthesized and their repair by human O6-alkylguanine-DNA alkyltransferase (hAGT) investigated. Duplexes containing 11 base-pairs with the ICLs in the center were assembled by automated DNA solid-phase synthesis using a cross-linked 2′-deoxyguanosine dimer phosphoramidite, prepared via a seven step synthesis which employed the Mitsunobu reaction to introduce the alkyl lesion at the O6 atom of guanine. Introduction of the four and seven carbon ICLs resulted in no change in duplex stability based on UV thermal denaturation experiments compared to a non-cross-linked control. Circular dichroism spectra of these ICL duplexes exhibited features of a B-form duplex, similar to the control, suggesting that these lesions induce little overall change in structure. The efficiency of repair by hAGT was examined and it was shown that hAGT repairs both ICL containing duplexes, with the heptyl ICL repaired more efficiently relative to the butyl cross-link. These results were reproducible with various hAGT mutants including one that contains a novel V148L mutation. The ICL duplexes displayed similar binding affinities to a C145S hAGT mutant compared to the unmodified duplex with the seven carbon containing ICLs displaying slightly higher binding. Experiments with CHO cells to investigate the sensitivity of these cells to busulfan and hepsulfam demonstrate that hAGT reduces the cytotoxicity of hepsulfam suggesting that the O6-2′-deoxyguanosine-alkyl-O6-2′-deoxyguanosine interstrand DNA cross-link may account for at least part of the cytotoxicity of this agent.

  已合成了5²-GNC-结构中具有4个和7个亚甲基连接的O6-2²-脱氧鸟苷-烷基-O6-2²-脱氧鸟苷DNA链间交联（ICL），并研究了人类O6-烷基鸟嘌呤-DNA烷基转移酶（hAGT）对其的修复。使用交联的2²-脱氧鸟苷二聚体亚磷酰胺，通过自动DNA固相合成组装了包含11个碱基对的ICL双链，该亚磷酰胺是通过七步合成制备的，其中采用Mitsunobu反应在鸟嘌呤的O6原子引入烷基损伤。与非交联对照相比，引入4个和7个碳的ICL不会改变双链的稳定性（基于UV热变性实验）。这些ICL双链的圆二色光谱表现出与对照类似的B型双链特征，表明这些损伤不会引起整体结构变化。研究了hAGT的修复效率，结果表明，hAGT可以修复两种ICL双链，其中庚基ICL的修复效率比丁基交联更高。这些结果在各种hAGT突变体中具有可重复性，包括一种含有新型V148L突变的突变体。与未修饰的双链相比，ICL双链与C
• Altering Residue 134 Confers an Increased Substrate Range of Alkylated Nucleosides to the E. coli OGT Protein
  作者：Nadia Schoonhoven、Derek O’Flaherty、Francis McManus、Lauralicia Sacre、Anne Noronha、M. Kornblatt、Christopher Wilds

  DOI：10.3390/molecules22111948

  日期：——

  for the removal of mutagenic alkyl adducts at the O⁶-atom of guanine and O⁴-atom of thymine. In the current study we set out to understand the role of the Ser134 residue in the Escherichia coli AGT variant OGT on substrate discrimination. The S134P mutation in OGT increased the ability of the protein to repair both O⁶-adducts of guanine and O⁴-adducts of thymine. However, the S134P variant was unable

  O 1-烷基鸟嘌呤-DNA烷基转移酶（AGT）是负责除去鸟嘌呤的O 3原子和胸腺嘧啶的O 3原子的诱变烷基加合物的蛋白质。在本研究中，我们着手了解大肠杆菌AGT变异体OGT中Ser134残基对底物识别的作用。OGT中的S134P突变增加了蛋白质修复鸟嘌呤的O 3-加合物和胸腺嘧啶的O 3-加合物的能力。然而，S134P变体不能像野生型OGT那样修复桥接DNA双链体中两个鸟嘌呤的O 5原子的链间交联（ICL）。当与人AGT蛋白（hAGT）比较时，S134P OGT变体显示出降低的对O 3-烷基化的活性，但对O 3-烷基化破坏逆转具有更大的底物范围。OGT残基134的作用与其在人类同源物中的功能相似，Pro140在赋予hAGT修复鸟嘌呤O⁶位置大加合物的能力方面至关重要。最后，展示了一种使用单链寡核苷酸底物在hAGT和模型核苷之间产生共价缀合物的方法。
• Preparation of Covalently Linked Complexes Between DNA and <i>O</i>⁶-Alkylguanine-DNA Alkyltransferase Using Interstrand Cross-Linked DNA
  作者：Francis P. McManus、Amardeep Khaira、Anne M. Noronha、Christopher J. Wilds

  DOI：10.1021/bc300553u

  日期：2013.2.20

  O-6-alkylguanine-DNA alkyltransferases (AGT) are responsible for the removal of alkylation at both the O-6 atom of guanine and O-4 atom of thymine. AGT homologues show vast substrate differences with respect to the size of the adduct and which alkylated atoms they can restore. The human AGT (hAGT) has poor capabilities for removal of methylation at the O-4 atom of thymidine, which is not the case in most homologues. No structural data are available to explain this poor hAGT repair. We prepared and characterized O(6)G-butylene-(OT)-T-4 (XLGT4) and O(6)G-heptylene-(OT)-T-4 (XLGT7) interstrand cross-linked (ICL) DNA as probes for hAGT and the Escherichia coli homologues, OCT and Ada-C, for the formation of DNA-ACT covalent complexes. XLGT7 reacted only with hAGT and did so with a cross-linking efficiency of 25%, while XLGT4 was inert to all ACT tested. The hAGT mediated repair of XLGT7 occurred slowly, on the order of hours as opposed to the repair of O-6-methyl-2'-deoxyguanosine which requires seconds. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the repair reaction revealed the formation of a covalent complex with an observed migration in accordance with a DNA-ACT complex. The identity of this covalent complex, as determined by mass spectrometry, was composed of a heptamethylene bridge between the O-4 atom of thymidine (in an 11-mer DNA strand) to residue Cys145 of hAGT. This procedure can be applied to produce well-defined covalent complexes between ACT with DNA.
• Synthesis, Characterization, and Repair of a Flexible<i>O</i>⁶-2′-Deoxyguanosine-alkylene-<i>O</i>⁶-2′-deoxyguanosine Intrastrand Cross-Link
  作者：Derek K. O'Flaherty、Christopher J. Wilds

  DOI：10.1002/chem.201501103

  日期：2015.7.13

  Oligonucleotides tethered by an alkylene linkage between the O6‐atoms of two consecutive 2′‐deoxyguanosines, which lack a phosphodiester linkage between these residues, have been synthesized as a model system of intrastrand cross‐linked (IaCL) DNA. UV thermal denaturation studies of duplexes formed between these butylene‐ and heptylene‐linked oligonucleotides with their complementary DNA sequences revealed about

  作为两个链内交联（IaCL）DNA的模型系统，已经合成了通过两个连续的2'-脱氧鸟苷的O 6原子之间的亚烷基键束缚的寡核苷酸，这些残基之间没有磷酸二酯键。这些丁烯和庚烯连接的寡核苷酸及其互补的DNA序列之间形成的双链体的UV热变性研究表明，相对于未修饰的双链体，其稳定性降低了约20°C。模型IaCL双链体的圆二色性光谱显示了B型DNA的特征，表明病变引起的总体扰动最小。研究了包含IaCL模型的双链体作为O 6的底物来自人和大肠杆菌（Ada-C和OGT）的-烷基鸟嘌呤DNA烷基转移酶（AGT）蛋白。发现人类AGT可以修复两种IaCL双链体，对庚烯和丁烯类似物的修复效率更高，这进一步增加了我们对该蛋白可以修复的底物的了解。