³²P-cyclic di-3′,5′-adenosine monophosphate | 28071-86-5

• 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: ³²P-cyclic di-3′,5′-adenosine monophosphate
• 英文别名: ³²P-c-di-AMP；³²P-labeled 3',5'-cAMP；[³²P]cAMP；[α-32P]-cAMP；(4aR,6R,7R,7aS)-6-(6-aminopurin-9-yl)-2-hydroxy-2-oxo-4a,6,7,7a-tetrahydro-4H-furo[3,2-d](232P)[1,3,2]dioxaphosphinin-7-ol
• CAS: 28071-86-5
• 化学式: C₁₀H₁₂N₅O₆P
• 分子量: 330.235
• InChiKey: IVOMOUWHDPKRLL-AJPCBXPMSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -2.6
• 重原子数：
  22
• 可旋转键数：
  1
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.5
• 拓扑面积：
  155
• 氢给体数：
  3
• 氢受体数：
  10

反应信息

• 作为产物：
  描述：

  [α-32P]ATP 在 diadenylate cyclase 、 sodium chloride 、 magnesium chloride 作用下， 以 aq. buffer 为溶剂， 生成 ³²P-cyclic di-3′,5′-adenosine monophosphate
  参考文献：
  名称：

  An HD-domain phosphodiesterase mediates cooperative hydrolysis of c-di-AMP to affect bacterial growth and virulence

  摘要：

  最近，小分子核苷酸环状二-3'，5'-腺苷酸单磷酸（c-di-AMP）作为一种细菌普遍存在的信号分子出现，对细菌生理和宿主-病原体相互作用都起着至关重要的作用。具有异常c-di-AMP水平的细菌突变体表现出生长和毒力缺陷，反映了调节c-di-AMP合成和降解对正常信号转导和适应环境变化的重要性。以前记录的磷酸二酯酶通过DHH-DHHA1结构域水解c-di-AMP，但并不是所有合成c-di-AMP的物种都具有这种酶。我们发现了一类以前未被认识的His-Asp结构域磷酸二酯酶，广泛分布于几个分类群中。此外，对于细菌病原体李斯特菌（Listeria monocytogenes），磷酸二酯酶突变体表现出宿主炎症增强、宿主细胞内生长缺陷以及在小鼠感染模型中明显的毒力减弱。

  DOI：

  10.1073/pnas.1416485112

文献信息

• Bis-Halogen-Anthraniloyl-Substituted Nucleoside 5′-Triphosphates as Potent and Selective Inhibitors of<i>Bordetella pertussis</i>Adenylyl Cyclase Toxin
  作者：Jens Geduhn、Stefan Dove、Yuequan Shen、Wei-Jen Tang、Burkhard König、Roland Seifert

  DOI：10.1124/jpet.110.174219

  日期：2011.1

  Whooping cough is caused by Bordetella pertussis and still constitutes one of the top five causes of death in young children, particularly in developing countries. The calmodulin-activated adenylyl cyclase (AC) toxin CyaA substantially contributes to disease development. Thus, potent and selective CyaA inhibitors would be valuable drugs for the treatment of whooping cough. However, it has been difficult to obtain potent CyaA inhibitors with selectivity relative to mammalian ACs. Selectivity is important for reducing potential toxic effects. In a previous study we serendipitously found that bis-methylanthraniloyl (bis-MANT)-IMP is a more potent CyaA inhibitor than MANT-IMP ( Mol Pharmacol 72: 526–535, 2007). These data prompted us to study the effects of a series of 32 bulky mono- and bis-anthraniloyl (ANT)-substituted nucleotides on CyaA and mammalian ACs. The novel nucleotides differentially inhibited CyaA and ACs 1, 2, and 5. Bis-ANT nucleotides inhibited CyaA competitively. Most strikingly, bis-Cl-ANT-ATP inhibited CyaA with a potency ≥100-fold higher than ACs 1, 2, and 5. In contrast to MANT-ATP, bis-MANT-ATP exhibited low intrinsic fluorescence, thereby substantially enhancing the signal-to noise ratio for the analysis of nucleotide binding to CyaA. The high sensitivity of the fluorescence assay revealed that bis-MANT-ATP binds to CyaA already in the absence of calmodulin. Molecular modeling showed that the catalytic site of CyaA is sufficiently spacious to accommodate both MANT substituents. Collectively, we have identified the first potent CyaA inhibitor with high selectivity relative to mammalian ACs. The fluorescence properties of bis-ANT nucleotides facilitate development of a high-throughput screening assay.

  百日咳由百日咳杆菌（Bordetella pertussis）引起，目前仍是导致幼儿死亡的五大原因之一，在发展中国家尤其如此。钙调蛋白激活的腺苷酸环化酶（AC）毒素 CyaA 在很大程度上导致了疾病的发生。因此，强效的选择性 CyaA 抑制剂将是治疗百日咳的重要药物。然而，要获得相对于哺乳动物 AC 具有选择性的强效 CyaA 抑制剂一直很困难。选择性对于减少潜在的毒性作用非常重要。在之前的一项研究中，我们偶然发现双甲基蒽酰（bis-MANT）-IMP 是一种比 MANT-IMP 更有效的 CyaA 抑制剂（《分子药理学》72：526-535，2007 年）。这些数据促使我们研究了一系列 32 种笨重的单蒽酰和双蒽酰（ANT）取代的核苷酸对 CyaA 和哺乳动物 AC 的影响。新型核苷酸对 CyaA 和 AC 1、2 和 5 有不同的抑制作用。双-ANT 核苷酸对 CyaA 有竞争性抑制作用。最引人注目的是，双Cl-ANT-ATP对CyaA的抑制作用比AC 1、2和5高出≥100倍。与 MANT-ATP 相反，双-MANT-ATP 显示出较低的本征荧光，从而大大提高了分析核苷酸与 CyaA 结合的信噪比。荧光检测的高灵敏度表明，在没有钙调蛋白的情况下，双-MANT-ATP 也能与 CyaA 结合。分子建模显示，CyaA 的催化位点足够宽敞，可以容纳两个 MANT 取代基。总之，我们发现了第一个对哺乳动物 AC 具有高选择性的强效 CyaA 抑制剂。双-ANT 核苷酸的荧光特性促进了高通量筛选试验的开发。
• Evolutionary Adaptation of the Essential tRNA Methyltransferase TrmD to the Signaling Molecule 3′,5′-cAMP in Bacteria
  作者：Yong Zhang、Rym Agrebi、Lauren E. Bellows、Jean-François Collet、Volkhard Kaever、Angelika Gründling

  DOI：10.1074/jbc.m116.758896

  日期：2017.1

  The nucleotide signaling molecule 3',5'-cyclic adenosine monophosphate (3',5'-cAMP) plays important physiological roles, ranging from carbon catabolite repression in bacteria to mediating the action of hormones in higher eukaryotes, including human. However, it remains unclear whether 3',5'-cAMP is universally present in the Firmicutes group of bacteria. We hypothesized that searching for proteins that bind 3',5'-cAMP might provide new insight into this question. Accordingly, we performed a genome-wide screen and identified the essential Staphylococcus aureus tRNA m(1)G37 methyltransferase enzyme TrmD, which is conserved in all three domains of life as a tight 3',5'-cAMP-binding protein. TrmD enzymes are known to use S-adenosyl-L-methionine (AdoMet) as substrate; we have shown that 3',5'-cAMP binds competitively with AdoMet to the S. aureus TrmD protein, indicating an overlapping binding site. However, the physiological relevance of this discovery remained unclear, as we were unable to identify a functional adenylate cyclase in S. aureus and only detected 2',3' -cAMP but not 3',5'-cAMP in cellular extracts. Interestingly, TrmD proteins from Escherichia coli and Mycobacterium tuberculosis, organisms known to synthesize 3',5'-cAMP, did not bind this signaling nucleotide. Comparative bioinformatics, mutagenesis, and biochemical analyses revealed that the highly conserved Tyr-86 residue in E. coli TrmD is essential to discriminate between 3',5'-cAMP and the native substrate AdoMet. Combined with a phylogenetic analysis, these results suggest that amino acids in the substrate binding pocket of TrmD underwent an adaptive evolution to accommodate the emergence of adenylate cyclases and thus the signaling molecule 3',5'-cAMP. Altogether this further indicates that S. aureus does not produce 3',5'-cAMP, which would otherwise competitively inhibit an essential enzyme.