1-[5-[[[[5-(6-氨基嘌呤-9-基)-3,4-二羟基-四氢呋喃-2-基]甲氧基-羟基-磷酰]氧基-羟基-磷酰]氧基甲基]-3,4-二羟基-四氢呋喃-2-基]吡啶-5-羧酸酯 | 6450-77-7

• 分子结构分类: 有机化合物 - 核苷、核苷酸和类似物 - （5'->5'）-二核苷酸
• 中文名称: 1-[5-[[[[5-(6-氨基嘌呤-9-基)-3,4-二羟基-四氢呋喃-2-基]甲氧基-羟基-磷酰]氧基-羟基-磷酰]氧基甲基]-3,4-二羟基-四氢呋喃-2-基]吡啶-5-羧酸酯
• 英文名称: nicotinic acid adenine dinucleotide
• 英文别名: NaAD；deamido-NAD zwitterion；[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2R,3S,4R,5R)-5-(3-carboxypyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate
• CAS: 6450-77-7
• 化学式: C₂₁H₂₆N₆O₁₅P₂
• 分子量: 664.416
• InChiKey: SENPVEZBRZQVST-HISDBWNOSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -4.6
• 重原子数：
  44
• 可旋转键数：
  10
• 环数：
  5.0
• sp3杂化的碳原子比例：
  0.48
• 拓扑面积：
  315
• 氢给体数：
  7
• 氢受体数：
  19

反应信息

• 作为反应物：
  描述：

  1-[5-[[[[5-(6-氨基嘌呤-9-基)-3,4-二羟基-四氢呋喃-2-基]甲氧基-羟基-磷酰]氧基-羟基-磷酰]氧基甲基]-3,4-二羟基-四氢呋喃-2-基]吡啶-5-羧酸酯 在 5’-三磷酸腺苷 (S)-2-[3]pyridyl-pyrrolidine-1-carboxylic acid amide 、 L-谷氨酰胺 、 NAD synthetase from Glycine max root nodule bacteroids 作用下， 以 水 为溶剂， 生成 β-烟酰胺腺嘌呤二核苷酸
  参考文献：
  名称：

  Formation of pyridine nucleotides under symbiotic and non-symbiotic conditions between soybean nodules and free-living rhizobia

  摘要：

  Enzymatic regulation of pyricline nucleotide formation, under symbiotic and non-symbiotic conditions, was analyzed using soybeans (Glycine max L. cv. 'Akisengoku') and rhizobia (Bradyrhizobia japonicum strain A1017), respectively. It was found that levels of pyridine nucleotides in bacteroids in root nodules were different from those in free-living cells of rhizobia. This difference was associated with differences in activities of enzymes involved in the pathway from L-tryptophan to NAD and NADP. That is, these activities were lower in bacteroids than in free-living bacteria and lower in the nodule cytosol than in root extracts. The optimum pH for NAD synthetase in bacteroids, was 9.0. Additionally, the optimum pH for ATP-nicotinamide mononucleotide (NMN) adenyltransferase, final step enzyme in NAD formation, was estimated to be 7.6. In the bacteroid fraction, the K-m of NAD synthetase (22 muM) was similar to 1/22 of that of ATP-NMN adenyltransferase (482 muM). Vmax values were estimated to be almost in the same order for both NAD synthetase and ATP-NMN adenyltransferase. This is the first report on the formation of pyridine nucleotides originating from L-tryptophan in bacteroids in soybean nodules and free-living bacteria. (C) 2002 Elsevier Science Ltd. All rights reserved.

  DOI：

  10.1016/s0031-9422(02)00364-3
• 作为产物：
  描述：

  alkaline earth salt of/the/ methylsulfuric acid 在 alkali 作用下， 生成 1-[5-[[[[5-(6-氨基嘌呤-9-基)-3,4-二羟基-四氢呋喃-2-基]甲氧基-羟基-磷酰]氧基-羟基-磷酰]氧基甲基]-3,4-二羟基-四氢呋喃-2-基]吡啶-5-羧酸酯
  参考文献：
  名称：

  Lamborg et al., Journal of Biological Chemistry, 1958, vol. 231, p. 685,692

  摘要：

  DOI：

文献信息

• A Universal, Fully Automated High Throughput Screening Assay for Pyrophosphate and Phosphate Release from Enzymatic Reactions
  作者：Scott Pegan、Yang Tian、Valerie Sershon、Andrew Mesecar

  DOI：10.2174/138620710790218203

  日期：2010.1.1

  The malachite green assay is often used for measuring the presence of inorganic mono-phosphate concentrations. Some studies have adapted this assay for use in monitoring enzymatic reactions and have suggested its potential use in high throughput screening (HTS). With the increasing availability of laboratory automation, some studies are starting to explore the possibility of conducting limited, semi-automated versions of the assay. Here we report the optimization and complete adaptation of the malachite green assay to a fully automated, HTS platform that can be performed unattended with standard, commercially available, automated liquid-handling systems. The assay is universal for the majority of enzymes that release phosphate or pyrophosphate. Moreover, the assay is fully scalable from smaller drug screening efforts (˜ 20,000 wells per day) to ultra-high throughput environments (˜ 200,000 wells per day). The assay uses cost-effective, commercially available reagents, and can be used to perform automated IC50 value and kinetic parameter determination. Finally, we demonstrate the utility of the assay via the initial, primary screening of 100,080 compounds against two target enzymes from Bacillus anthracis, O-succinylbenzoyl-CoA synthetase and nicotinate mononucleotide adenylyltransferase.

  孔雀石绿测定法通常用于测量无机单磷酸盐浓度的存在。一些研究已将这种检测方法改用于监测酶反应，并建议将其用于高通量筛选（HTS）。随着实验室自动化程度的不断提高，一些研究开始探索进行有限的半自动化测定的可能性。在此，我们报告了孔雀石绿测定法的优化和完全适应全自动 HTS 平台的情况，该平台可在无人看管的情况下使用标准的商用自动液体处理系统进行测定。该检测方法适用于大多数释放磷酸盐或焦磷酸盐的酶。此外，该检测方法完全可以从较小的药物筛选工作（˜ 20,000 孔/天）扩展到超高通量环境（˜ 200,000 孔/天）。该检测方法使用成本效益高、市场上可买到的试剂，可用于自动测定 IC50 值和动力学参数。最后，我们通过对 100,080 种化合物针对炭疽芽孢杆菌的两种目标酶（O-琥珀酰苯甲酰-CoA 合成酶和烟酸单核苷酸腺苷酸转移酶）进行初步初筛，证明了该检测方法的实用性。
• Nickel-pincer cofactor biosynthesis involves LarB-catalyzed pyridinium carboxylation and LarE-dependent sacrificial sulfur insertion
  作者：Benoît Desguin、Patrice Soumillion、Pascal Hols、Robert P. Hausinger

  DOI：10.1073/pnas.1600486113

  日期：2016.5.17

  Nicotinic acid is a precursor of the ubiquitous cofactors nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP). Previous studies revealed that nicotinic acid is required for lactate racemization, an enzymatic activity that enables microorganisms to produce or use both isomers of lactate. Lactate racemase (Lar) was shown to harbor a nicotinic acid-derived cofactor that coordinates a nickel ion, forming a pincer complex. The biosynthesis of this cofactor requires three accessory proteins: LarB, LarC, and LarE. Here, we describe this biosynthetic pathway by showing that LarB carboxylates and hydrolyzes the NAD precursor nicotinic acid adenine dinucleotide (NaAD) and LarE sacrificially inserts two sulfur atoms into the product of LarB. Finally, LarC inserts the nickel ion to form the mature cofactor.

  烟酸是普遍存在的辅因子烟酰胺腺嘌呤二核苷酸（NAD）和烟酰胺腺嘌呤二核苷酸磷酸（NADP）的前体。先前的研究表明，烟酸对乳酸拉曼化是必需的，这是一种酶活性，使微生物能够产生或使用乳酸的两种异构体。已经证明乳酸拉曼酶（Lar）具有一种烟酸衍生的辅因子，它协调一个镍离子，形成一个钳形配合物。这种辅因子的生物合成需要三种辅助蛋白：LarB、LarC和LarE。在这里，我们通过展示LarB羧化和水解NAD前体烟酸腺嘌呤二核苷酸（NaAD），以及LarE牺牲性地将两个硫原子插入LarB产物中来描述这种生物合成途径。最后，LarC插入镍离子形成成熟的辅因子。

• Serlupi-Crescenzi; Ballio, Gazzetta Chimica Italiana, 1958, vol. 88, p. 320,325
  作者：Serlupi-Crescenzi、Ballio

  DOI：——

  日期：——