cytidine monophospho-N-acetylneuraminic acid

• 分子结构分类: 有机化合物 - 核苷、核苷酸和类似物 - 嘧啶核苷酸
• 英文名称: cytidine monophospho-N-acetylneuraminic acid
• 英文别名: cytidine monophosphate N-acetylneuraminic acid；(2R,4S,5R,6R)-2-[[(2R,3S,4R,5R)-5-(4-amino-2-oxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-4-hydroxy-5-(1-oxidoethylideneamino)-6-[(1R,2R)-1,2,3-trihydroxypropyl]oxane-2-carboxylate
• 化学式: C₂₀H₂₉N₄O₁₆P
• 分子量: 612.441
• InChiKey: TXCIAUNLDRJGJZ-BILDWYJOSA-L

计算性质

• 辛醇/水分配系数(LogP)：
  -6.2
• 重原子数：
  41
• 可旋转键数：
  10
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.7
• 拓扑面积：
  326
• 氢给体数：
  8
• 氢受体数：
  16

反应信息

• 作为反应物：
  描述：

  cytidine monophospho-N-acetylneuraminic acid 、 (4,5-dimethoxy-2-nitrophenyl)methyl β-D-galactopyranosyl-(1->4)-β-D-glucopyranoside 在 α-2,3-sialyltransferase disodium ethylenediaminetetraacetic acid 、 sodium chloride 、 Triton X-100 作用下， 以 水 为溶剂， 反应 48.0h， 以70%的产率得到4,5-dimethoxy-2-nitrobenzyl O-5-acetamido-3,5-dideoxy-D-glycero-α-D-galacto-2-nonulopyranosylonic acid-(2->3)-O-β-D-galactopyranosyl-(1->4)-β-D-glucopyranoside sodium salt
  参考文献：
  名称：

  Synthesis and Characterization of an Anomeric Sulfur Analogue of CMP-Sialic Acid

  摘要：

  alpha-2,3-Sialyltransferase catalyzes the transfer of sialic acid from CMP-sialic acid (1) to a lactose acceptor. An analogue of 1 was synthesized in which the anomeric oxygen atom was replaced with a sulfur atom (1S). The key step in the synthesis of 1S was a tetrazole-promoted coupling of a cytidine-5'-phosphoramidite with a glycosyl thiol of a protected sialic acid. Compounds 1 and 1S were characterized for their activity in a sialyl transfer assay. The rate of solvolysis in aqueous buffer of analogue 1S was 50-fold slower than that of 1. Analogue 1S was found to be substrate for alpha-2,3-sialyltransferase. The K-m of 1S was just 3-fold higher than that of 1, while the k(cat) of 1S was 2 orders of magnitude lower compared to 1.

  DOI：

  10.1021/jo000646+
• 作为产物：
  描述：

  allyl 2',3'-O,N⁴-triacetylcytidin-5'-yl methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-β-D-glycero-D-galacto-2-nonulopyranosid-2''-yl phosphite 在 甲醇 、 叔丁基过氧化氢 、 sodium methylate 、 三乙胺 作用下， 生成 cytidine monophospho-N-acetylneuraminic acid
  参考文献：
  名称：

  Enzyme-Catalyzed Synthesis of Oligosaccharides That Contain Functionalized Sialic Acids

  摘要：

  DOI：

  10.1021/ja963894p

文献信息

• Synthesis of Bisubstrate and Donor Analogues of Sialyltransferase and Their Inhibitory Activities
  作者：Masayuki Izumi、Katsuhiro Wada、Hideya Yuasa、Hironobu Hashimoto

  DOI：10.1021/jo0512608

  日期：2005.10.1

  Sialyltransferases (STs) are involved in the biosynthesis of glycoconjugates with important biological activities. Most STs utilize cytidine-5‘-monophospho-N-acetylneuraminic acid (CMP−Neu5Ac) as a common donor substrate. A bisubstrate analogue containing the donor substrate (CMP−Neu5Ac mimic) and the acceptor substrate (galactose) was synthesized. Four donor analogues having the partial structure

  唾液酸转移酶（STs）参与具有重要生物学活性的糖缀合物的生物合成。大多数ST利用胞苷-5'-单磷酸-N-乙酰神经氨酸（CMP-Neu5Ac）作为共同的供体底物。合成了包含供体底物（CMP-Neu5Ac模拟物）和受体底物（半乳糖）的双底物类似物。具有双底物类似物的部分结构的四个供体类似物也被合成以支持对结构-活性关系的研究。每个类似物都包含一个亚乙基，代替CMP-Neu5Ac的外环异头氧。双底物类似物仅对大鼠重组α-2,3-和α-2,6-ST表现出微弱的抑制活性（IC 50= 1.3，2.4 mM）。Neu5Ac部分的C-1羧酸盐转化为羧酰胺，羟甲基或磷酸亚甲基酯均导致抑制活性降低。在合成的类似物中，胞苷5'-唾液酸乙基膦酸酯（4）是对抗大鼠重组α-2,3-和α-2,6-ST的最有效抑制剂（IC 50 = 0.047，0.34 mM）。
• Flexibility of Substrate Binding of Cytosine-5′-Monophosphate-N-Acetylneuraminate Synthetase (CMP-Sialate Synthetase) from Neisseria meningitidis: An Enabling Catalyst for the Synthesis of Neo-sialoconjugates
  作者：Ning He、Dong Yi、Wolf-Dieter Fessner

  DOI：10.1002/adsc.201100412

  日期：2011.9

  sensitive and reliable quantification of cytosine‐5′‐monophosphate‐acetylneuraminate synthetase (CMP‐sialate synthetase, CSS) activity based on the pH change of a released proton equivalent upon nucleotide activation of Neu5Ac or related analogues. Using this method, steady‐state kinetic data of Neisseria meningitidis CSS were determined for cytosine‐5′‐triphosphate (CTP), N‐acetylneuraminic acid (Neu5Ac)

  我们已经成功建立了一个简单的连续比色测定法，可基于Neu5Ac或NPS核苷酸激活后释放的质子当量的pH值变化，对胞嘧啶5'-单磷酸酯-乙酰神经氨酸合成酶（CMP-唾液酸合成酶，CSS）活性进行灵敏可靠的定量分析。相关类似物。使用此方法，确定了脑膜炎奈瑟氏球菌CSS的稳态动力学数据，包括胞嘧啶5'-三磷酸（CTP），N-乙酰神经氨酸（Neu5Ac）和唾液酸的11种结构变异，包括主链截短，脱氨基，差向异构化，和几个N酰基修饰。这些数据进一步证明了脑膜炎奈瑟氏球菌具有非同寻常的多功能性CSS，用于一般访问包含非天然唾液酸类似物的唾液缀合物。引人注目的是，该测定法可以覆盖范围广泛的底物参数，这些参数涵盖了超过3个数量级的K M和k cat测量。借助基于X射线晶体结构数据构建的结构模型，动力学数据可用于解释Neu5Ac及其类似物在底物结合中的潜在蛋白质贡献。使用脑膜炎奈瑟氏球菌CSS和来自Photobacter的新型α2
• Comparison of the Enzymatic Properties of Mouse  -Galactoside  2,6-Sialyltransferases, ST6Gal I and II
  作者：S. Takashima

  DOI：10.1093/jb/mvg142

  日期：2003.8.1

  The cDNA encoding a second type of mouse β-galactoside α2,6-sialyltransferase (ST6Gal II) was cloned and characterized. The sequence of mouse ST6Gal II encoded a protein of 524 amino acids and showed 77.1% amino acid sequence identity with human ST6Gal II. Recombinant ST6Gal II exhibited α2,6-sialyltransferase activity toward oligosaccharides that have the Galβ1,4GlcNAc sequence at the nonreducing end of their carbohydrate groups, but it exhibited relatively low and no activity toward some glycoproteins and glycolipids, respectively. On the other hand, ST6Gal I, which has been known as the sole member of the ST6Gal-family for more than ten years, exhibited broad substrate specificity toward oligosaccharides, glycoproteins, and a glycolipid, paragloboside. The ST6Gal II gene was mainly expressed in brain and embryo, whereas the ST6Gal I gene was ubiquitously expressed, and its expression levels were higher than those of the ST6Gal II gene. The ST6Gal II gene is located on chromosome 17 and spans over 70 kb of mouse genomic DNA consisting of at least 6 exons. The ST6Gal II gene has a similar genomic structure to the ST6Gal I gene. In this paper, we have shown that ST6Gal II is a counterpart of ST6Gal I.

  克隆并鉴定了编码小鼠第二种β-半乳糖苷α2,6-糖基转移酶（ST6Gal II）的cDNA。小鼠 ST6Gal II 的序列编码了一个 524 个氨基酸的蛋白质，与人 ST6Gal II 的氨基酸序列相同度为 77.1%。重组 ST6Gal II 对碳水化合物基团非还原端具有 Galβ1,4GlcNAc序列的寡糖具有α2,6-苷基转移酶活性，但对一些糖蛋白和糖脂的活性相对较低，甚至没有活性。另一方面，ST6Gal I 作为 ST6Gal 家族的唯一成员已有十多年的历史，它对寡糖、糖蛋白和一种糖脂--paragloboside 具有广泛的底物特异性。ST6Gal II 基因主要在脑和胚胎中表达，而 ST6Gal I 基因则普遍表达，其表达水平高于 ST6Gal II 基因。ST6Gal II 基因位于第 17 号染色体上，在小鼠基因组 DNA 中的跨度超过 70 kb，由至少 6 个外显子组成。ST6Gal II 基因的基因组结构与 ST6Gal I 基因相似。本文表明，ST6Gal II 是 ST6Gal I 的对应基因。
• Identification and functional expression of a second human β-galactoside α2,6-sialyltransferase, ST6Gal II
  作者：Marie-Ange Krzewinski-Recchi、Sylvain Julien、Sylvie Juliant、Mélanie Teintenier-Lelièvre、Bénédicte Samyn-Petit、Maria-Dolores Montiel、Anne-Marie Mir、Martine Cerutti、Anne Harduin-Lepers、Philippe Delannoy

  DOI：10.1046/j.1432-1033.2003.03458.x

  日期：2003.3

  blast analysis of the human and mouse genome sequence databases using the sequence of the human CMP‐sialic acid:β‐galactoside α‐2,6‐sialyltransferase cDNA (hST6Gal I, EC2.4.99.1) as a probe allowed us to identify a putative sialyltransferase gene on chromosome 2. The sequence of the corresponding cDNA was also found as an expressed sequence tag of human brain. This gene contained a 1590 bp open reading frame divided in five exons and the deduced amino‐acid sequence didn't correspond to any sialyltransferase already known in other species. Multiple sequence alignment and subsequent phylogenic analysis showed that this new enzyme belonged to the ST6Gal subfamily and shared 48% identity with hST6Gal‐I. Consequently, we named this new sialyltransferase ST6Gal II. A construction in pFlag vector transfected in COS‐7 cells gave raise to a soluble active form of ST6Gal II. Enzymatic assays indicate that the best acceptor substrate of ST6Gal II was the free disaccharide Galβ1–4GlcNAc structure whereas ST6Gal I preferred Galβ1–4GlcNAc‐R disaccharide sequence linked to a protein. The α2,6‐linkage was confirmed by the increase of Sambucus nigra agglutinin‐lectin binding to the cell surface of CHO transfected with the cDNA encoding ST6Gal II and by specific sialidases treatment. In addition, the ST6Gal II gene showed a very tissue specific pattern of expression because it was found essentially in brain whereas ST6Gal I gene is ubiquitously expressed.

  通过对人类和小鼠基因组序列数据库进行BLAST分析，并使用人类CMP-唾液酸-β-半乳糖苷α-2,6-唾液酸转移酶cDNA（hST6Gal I，EC2.4.99.1）的序列作为探针，我们鉴定出一个位于2号染色体上的潜在唾液酸转移酶基因。相应的cDNA序列也被发现是人脑的表达序列标签（EST）。该基因包含一个1590个碱基对的开放阅读框，分为5个外显子。推导出的氨基酸序列与目前已知的其他物种的唾液酸转移酶无对应关系。 多序列比对和随后的系统发育分析表明，这种新酶属于ST6Gal亚家族，并与hST6Gal-I共享了48%的序列一致性。因此，我们将这种新的唾液酸转移酶命名为ST6Gal II。使用pFLAg载体构建并转染COS-7细胞后，获得了ST6Gal II的可溶性活性形式。酶活性测定表明，ST6Gal II的最佳受体底物是游离的二糖结构Galβ1–4GlcNAc，而hST6Gal-I则偏好与蛋白质相连的Galβ1–4GlcNAc-R二糖序列。 通过增加Sambucus nigra凝集素- lECtin与转染了编码ST6Gal II的cDNA的CHO细胞表面的结合量，以及通过特异性神经氨酸酶处理，证实了α2,6连接的存在。此外，ST6Gal II基因显示出非常特异的组织表达模式，主要在大脑中发现，而ST6Gal I基因则是广泛表达的。
• Universal phosphatase-coupled glycosyltransferase assay
  作者：Z. L. Wu、C. M. Ethen、B. Prather、M. Machacek、W. Jiang

  DOI：10.1093/glycob/cwq187

  日期：2011.6.1

  A nonradioactive glycosyltransferase assay is described here. This method takes advantage of specific phosphatases that can be added into glycosyltransferase reactions to quantitatively release inorganic phosphate from the leaving groups of glycosyltransferase reactions. The released phosphate group is then detected using colorimetric malachite-based reagents. Because the amount of phosphate released is directly proportional to the sugar molecule transferred in a glycosyltransferase reaction, this method can be used to obtain accurate kinetic parameters of the glycosyltransferase. The assay can be performed in multiwell plates and quantitated by a plate reader, thus making it amenable to high-throughput screening. It has been successfully applied to all glycosyltransferases available to us, including glucosyltransferases, N-acetylglucosaminyltransferases, N-acetylgalactosyltransferases, galactosyltransferases, fucosyltransferases and sialyltransferases. As examples, we first assayed Clostridium difficile toxin B, a protein O-glucosyltransferase that specifically monoglucosylates and inactivates Rho family small GTPases; we then showed that human KTELC1, a homolog of Rumi from Drosophila, was able to hydrolyze UDP-Glc; and finally, we measured the kinetic parameters of human sialyltransferase ST6GAL1.

  这里介绍的是一种非放射性糖基转移酶测定法。该方法利用可加入糖基转移酶反应的特异性磷酸酶，定量释放糖基转移酶反应离去基团中的无机磷酸盐。然后使用孔雀石基比色试剂检测释放的磷酸基团。由于释放的磷酸量与糖基转移酶反应中转移的糖分子成正比，因此这种方法可用于获得糖基转移酶的精确动力学参数。该测定可在多孔板中进行，并通过平板阅读器进行定量，因此适合于高通量筛选。它已成功应用于我们现有的所有糖基转移酶，包括葡萄糖基转移酶、N-乙酰葡萄糖氨基转移酶、N-乙酰半乳糖基转移酶、半乳糖基转移酶、岩藻糖基转移酶和硅丙基转移酶。例如，我们首先检测了艰难梭菌毒素B，它是一种蛋白质O-葡糖基转移酶，能特异性地使Rho家族小GTP酶单葡糖基化并失活；然后，我们发现果蝇Rumi的同源物人类KTELC1能水解UDP-Glc；最后，我们测量了人类硅氨酰转移酶ST6GAL1的动力学参数。