¹³C₁₀/¹⁵N₅-ATP

• 分子结构分类: 有机化合物 - 核苷、核苷酸和类似物 - 嘌呤核苷酸
• 英文名称: ¹³C₁₀/¹⁵N₅-ATP
• 英文别名: ((2R,3S,4R,5R)-5-(6-(Amino-15N)-9H-purin-9-yl-13C5-15N4)-3,4-dihydroxytetrahydrofuran-2-yl-2,3,4,5-13C4)methyl-13C tetrahydrogen triphosphate；[[(2R,3S,4R,5R)-5-(6-(15N)azanylpurin-9-yl)-3,4-dihydroxy(2,3,4,5-13C4)oxolan-2-yl](113C)methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate
• 化学式: C₁₀H₁₆N₅O₁₃P₃
• 分子量: 522.041
• InChiKey: ZKHQWZAMYRWXGA-IADJQYSOSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -5.7
• 重原子数：
  31
• 可旋转键数：
  8
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.5
• 拓扑面积：
  279
• 氢给体数：
  7
• 氢受体数：
  17

反应信息

• 作为反应物：
  描述：

  ¹³C₁₀/¹⁵N₅-ATP 在 ribonucleotide triphosphate reductase 、 重水 作用下， 生成 [[(2R,3S,4R,5R)-5-(6-(15N)azanylpurin-9-yl)-4-deuterio-3-hydroxy(2,3,4,5-13C4)oxolan-2-yl](113C)methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate
  参考文献：
  名称：

  Determination of DNA Structure in Solution:  Enzymatic Deuteration of the Ribose 2‘ Carbon

  摘要：

  An enzymatic solution to the problem of obtaining 13C/15N-labeled nucleotides that are deuterated uniquely at the H2' ' position within the ribose ring is presented. Selective deuteration occurs with an overall yield of >80%. The deuteron at the H2' ' position allows measurement of the scalar and residual dipolar couplings for the bond vectors attached to the C2' carbon of each ribose sugar. These data allow the accurate determination of sugar conformation. Interesting DNA double helices of 2-3 turns are now within the reach of solution NMR spectroscopy. As an example, these labeled nucleotides are incorporated uniquely at positions 6-14 in a 20-bp DNA sequence containing the adenovirus major late promoter.

  DOI：

  10.1021/ja026678r
• 作为产物：
  描述：

  ^[13C,15N]AMP 在 calcium chloride 、 apyrase 作用下， 以 aq. buffer 为溶剂， 反应 0.5h， 生成 ¹³C₁₀/¹⁵N₅-ATP
  参考文献：
  名称：

  LC-MS / MS稳定同位素标记的内标用于磷酸化代谢产物定量的合成及应用

  摘要：

  液相色谱与串联质谱联用（LC-MS / MS）是测量代谢产物的高度特异性和灵敏的技术。但是，组织提取物中的共洗脱成分可能会干扰LC和MS / MS相界面处的电离，从而导致代谢物浓度低估或高估。掺入已知量的稳定同位素标记的内标（SIL-IS）的样品可使相应代谢物的测量值针对此类基质效应进行校正。我们描述了选择合适的SIL-IS的标准，并报告了六种SIL-IS的酶法合成和纯化，分别用于己糖，戊糖和三糖磷酸酯，UDP葡萄糖和单磷酸腺苷（AMP）。连同用于其他7种代谢物的市售SIL-IS，莱茵衣藻，大肠杆菌和面包酵母（Saccharomyces cerevisiae）。除少数例外，使用SIL-IS加标显着提高了在多种萃取物稀释液中基于LC-MS / MS的代谢物测量结果的可重复性，表明该方法可有效校正基质效应。通过使用SIL-IS校正基质效应，LC-MS / MS为可靠地确定各种生物体中的光合和呼吸中间产物提供了前所未有的范围。

  DOI：

  10.1021/acs.analchem.5b01387

文献信息

• Synthesis and application of isotopically labeled flavin nucleotides
  作者：Tatiana V. Mishanina、Amnon Kohen

  DOI：10.1002/jlcr.3313

  日期：2015.7

  Flavin nucleotides, i.e. flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are utilized as prosthetic groups and/or substrates by a myriad of proteins, ranging from metabolic enzymes to light receptors. Isotopically labeled flavins have served as invaluable tools in probing the structure and function of these flavoproteins. Here we present an enzymatic synthesis of several radio- and stable-isotope labeled flavin nucleotides from commercially available labeled riboflavin and ATP. The synthetic procedure employs a bifunctional enzyme, Corynebacterium ammoniagenes FAD synthetase, that sequentially converts riboflavin to FMN and then to FAD. The final flavin product (FMN or FAD) is controlled by the concentration of ATP in the reaction. Utility of the synthesized labeled FAD cofactors is demonstrated in flavin-dependent thymidylate synthase. The described synthetic approach can be easily applied to the production of flavin nucleotide analogues from riboflavin precursors.

  黄素核苷酸，即黄素单核苷酸（FMN）和黄素腺嘌呤二核苷酸（FAD），被广泛用作从代谢酶到光感受器等多种蛋白质的辅基和/或底物。同位素标记的黄素是探究这些黄素蛋白结构与功能的宝贵工具。本文介绍了一种酶促合成多种放射性和稳定同位素标记的黄素核苷酸的方法，该方法利用商业可得的标记核黄素和ATP作为原料。合成过程采用一种双功能酶——戈登链霉菌FAD合成酶，该酶依次将核黄素转化为FMN，再转化为FAD。最终的黄素产物（FMN或FAD）由反应中的ATP浓度控制。合成标记的FAD辅因子在黄素依赖型胸苷酸合酶中的应用效果得到验证。所描述的合成方法可轻松应用于从核黄素前体生产黄素核苷酸类似物。
• Radical SAM Enzyme HydE Generates Adenosylated Fe(I) Intermediates En Route to the [FeFe]-Hydrogenase Catalytic H-Cluster
  作者：Lizhi Tao、Scott A. Pattenaude、Sumedh Joshi、Tadhg P. Begley、Thomas B. Rauchfuss、R. David Britt

  DOI：10.1021/jacs.0c03802

  日期：2020.6.17

  the substrate of the radical SAM enzyme HydE, with the generated 5'-deoxyadenosyl radical attacking the cysteine S to form a C5'-S bond concomitant with reduction of the central low-spin Fe(II) to the Fe(I) oxidation state. This leads to the cleavage of the cysteine C3-S bond, producing a mononuclear [FeI(CO)2(CN)S] species that serves as the precursor to the dinuclear Fe(I)Fe(I) center of [2Fe]H-subcluster

  [FeFe]-氢化酶的 H-簇由 [4Fe-4S]H-亚簇组成，通过半胱氨酰桥连接到独特的有机金属 [2Fe]H-亚簇，指定为质子和分子氢之间相互转化的位点。这个 [2Fe]H 亚簇由一组 Fe-S 成熟酶 HydG、HydE 和 HydF 组装而成。在这里，我们表明 HydG 产物 [FeII(Cys)(CO)2(CN)] 合成子是自由基 SAM 酶 HydE 的底物，生成的 5'-脱氧腺苷自由基攻击半胱氨酸 S 形成 C5'-S键伴随着中心低自旋 Fe(II) 还原为 Fe(I) 氧化态。这导致半胱氨酸 C3-S 键断裂，产生单核 [FeI(CO)2(CN)S] 物质，作为 [2Fe]H 双核 Fe(I)Fe(I) 中心的前体- 子集群。
• Synthesis and Use of Stable-Isotope-Labeled Internal Standards for Quantification of Phosphorylated Metabolites by LC–MS/MS
  作者：Stéphanie Arrivault、Manuela Guenther、Stephen C. Fry、Maximilian M. F. F. Fuenfgeld、Daniel Veyel、Tabea Mettler-Altmann、Mark Stitt、John E. Lunn

  DOI：10.1021/acs.analchem.5b01387

  日期：2015.7.7

  tandem mass spectrometry (LC–MS/MS) is a highly specific and sensitive technique for measuring metabolites. However, coeluting components in tissue extracts can interfere with ionization at the interface of the LC and MS/MS phases, causing under- or overestimation of metabolite concentrations. Spiking of samples with known amounts of stable-isotope-labeled internal standards (SIL-IS) allows measurements

  液相色谱与串联质谱联用（LC-MS / MS）是测量代谢产物的高度特异性和灵敏的技术。但是，组织提取物中的共洗脱成分可能会干扰LC和MS / MS相界面处的电离，从而导致代谢物浓度低估或高估。掺入已知量的稳定同位素标记的内标（SIL-IS）的样品可使相应代谢物的测量值针对此类基质效应进行校正。我们描述了选择合适的SIL-IS的标准，并报告了六种SIL-IS的酶法合成和纯化，分别用于己糖，戊糖和三糖磷酸酯，UDP葡萄糖和单磷酸腺苷（AMP）。连同用于其他7种代谢物的市售SIL-IS，莱茵衣藻，大肠杆菌和面包酵母（Saccharomyces cerevisiae）。除少数例外，使用SIL-IS加标显着提高了在多种萃取物稀释液中基于LC-MS / MS的代谢物测量结果的可重复性，表明该方法可有效校正基质效应。通过使用SIL-IS校正基质效应，LC-MS / MS为可靠地确定各种生物体中的光合和呼吸中间产物提供了前所未有的范围。
• Determination of DNA Structure in Solution:  Enzymatic Deuteration of the Ribose 2‘ Carbon
  作者：Douglas MacDonald、Ponzy Lu

  DOI：10.1021/ja026678r

  日期：2002.8.1

  An enzymatic solution to the problem of obtaining 13C/15N-labeled nucleotides that are deuterated uniquely at the H2' ' position within the ribose ring is presented. Selective deuteration occurs with an overall yield of >80%. The deuteron at the H2' ' position allows measurement of the scalar and residual dipolar couplings for the bond vectors attached to the C2' carbon of each ribose sugar. These data allow the accurate determination of sugar conformation. Interesting DNA double helices of 2-3 turns are now within the reach of solution NMR spectroscopy. As an example, these labeled nucleotides are incorporated uniquely at positions 6-14 in a 20-bp DNA sequence containing the adenovirus major late promoter.