phosphoenolpyruvate | 67533-07-7

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 有机磷酸及其衍生物
• 英文名称: phosphoenolpyruvate
• 英文别名: PEP；Phosphonatoenolpyruvate；2-phosphonatooxyprop-2-enoate
• CAS: 67533-07-7
• 化学式: C₃H₂O₆P
• 分子量: 165.019
• InChiKey: DTBNBXWJWCWCIK-UHFFFAOYSA-K

计算性质

• 辛醇/水分配系数(LogP)：
  -0.6
• 重原子数：
  10
• 可旋转键数：
  1
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.0
• 拓扑面积：
  113
• 氢给体数：
  0
• 氢受体数：
  6

反应信息

• 作为反应物：
  描述：

  phosphoenolpyruvate 以 水 为溶剂， 生成 piruvate
  参考文献：
  名称：

  腺苷3',5'-(环)磷酸(aq)和磷酸烯醇式丙酮酸(aq)水解反应的热力学；3',5'-(环状)磷酸酯(aq)和磷酸烯醇式丙酮酸(aq)的标准摩尔形成特性

  摘要：

  摘要 已经测量了生化反应 {cAMP(aq) + H2O(l)=AMP(aq)} 和 {PEP(aq) + H2O(l)=丙酮酸盐(aq) + 磷酸盐的摩尔量热焓变 ΔrHm(cal) (aq)}。该反应分别由磷酸二酯酶 3',5'-环核苷酸和碱性磷酸酶催化。通过使用化学平衡模型来分析结果以获得各个参考反应{cAMP-(aq) + H2O(l)=HAMP-(aq)}和{PEP3-(aq)的标准摩尔反应焓值ΔrHm∘ ) + H2O(l)=丙酮酸-(aq) + HPO2-4(aq)}。反应 {ATP(aq)=cAMP(aq) + 焦磷酸盐(aq)}, {ATP(aq) + 丙酮酸(aq)=ADP(aq) + PEP(aq)} 的表观平衡常数 K' 的文献值，{ATP(aq) + pyruvate(aq) + 磷酸盐(aq)=AMP(aq) + PEP(aq) +焦磷酸盐(aq)}也使用化学平衡模型进行分析。这些计算产生了平衡常数

  DOI：

  10.1016/j.jct.2003.08.002
• 作为产物：
  描述：

  2-氧代丁二酸 在 pyruvate kinase 、 recombinant rat cytosolic phosphoenolpyruvate carboxykinase 、 二磷酸腺苷 、 Guanosine 5'-triphosphate 、 还原型辅酶Ⅰ 、 magnesium chloride 、 manganese(ll) chloride 、 L-lactate dehydrogenase 、 1,4-二巯基-2,3-丁二醇 作用下， 以 水 为溶剂， 生成 phosphoenolpyruvate
  参考文献：
  名称：

  Increasing the Conformational Entropy of the Ω-Loop Lid Domain in Phosphoenolpyruvate Carboxykinase Impairs Catalysis and Decreases Catalytic Fidelity,

  摘要：

  Many studies have shown that the dynamic motions of individual protein segments can play an important role in enzyme function. Recent structural studies of the gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK) demonstrate that PEPCK contains a 10-residue Omega-loop domain that acts as an active site lid. On the basis of these structural studies, we have previously proposed a model for the mechanism of PEPCK catalysis in which the conformation of this mobile lid domain is energetically coupled to ligand binding, resulting in the closed conformation of the lid, necessary for correct substrate positioning, becoming more energetically favorable as ligands associate with the enzyme. Here we test this model by introducing a point mutation (A467G) into the center of the Omega-loop lid that is designed to increase the entropic penalty for lid closure. Structural and kinetic characterization of this mutant enzyme demonstrates that the mutation has decreased the favorability of the enzyme adapting the closed lid conformation. As a consequence of this shift in the equilibrium defining the conformation of the active site lid, the enzyme's ability to stabilize the reaction intermediate is weakened, resulting in catalytic defect. This stabilization is initially surprising, as the lid domain makes no direct contacts with the etiolate intermediate formed during the reaction. Furthermore, during the conversion of OAA to PEP, the destabilization of the lid-closed conformation results in the reaction becoming decoupled as the etiolate intermediate is protonated rather than phosphorylated, resulting in the formation of pyruvate. Taken together, the structural and kinetic characterization of A467G-PEPCK supports our model of the role of the active site lid in catalytic function and demonstrates that the shift in the lowest-energy conformation between open and closed lid states is a function of the free energy available to the enzyme through ligand binding and the entropic penalty for ordering of the 10-residue Omega-loop lid domain.

  DOI：

  10.1021/bi100399e
• 作为试剂：
  描述：

  阿米卡星 在 pyruvate kinase 、 recombinant Enterococcus faecium aminoglycoside 2''-phosphotransferase type IIa N₁₉₆D/D₂₆₈N mutant 、 potassium chloride 、 phosphoenolpyruvate 、 5’-三磷酸腺苷 、 还原型辅酶Ⅰ 、 magnesium chloride 、 L-lactate dehydrogenase 作用下， 生成 [(2S,3R,4S,5S,6R)-4-amino-2-[(1S,2S,3R,4S,6R)-4-amino-6-[[(2S)-4-amino-2-hydroxybutanoyl]amino]-3-[(2R,3R,4S,5S,6R)-6-(aminomethyl)-3,4,5-trihydroxyoxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-3-yl] dihydrogen phosphate
  参考文献：
  名称：

  Mutant APH(2″)-IIa Enzymes with Increased Activity against Amikacin and Isepamicin

  摘要：

  摘要 通过随机 PCR 诱变氨基糖苷 2″-IIa 磷酸转移酶基因的定向进化产生了 R92H/D268N 和 N196D/D268N 突变体酶，导致对阿米卡星和异帕米星的耐药性增强，但对其他氨基糖苷类抗生素的耐药性没有增强。突变型磷酸转移酶对异帕米星的活性增加是由于其对阿米卡星和异帕米星的耐药性降低。 K m 值降低的结果，而阿米卡星催化效率的提高则是 K m K m 值降低和抗生素周转率增加的结果。R92H、D268N 和 D268N 单个氨基酸置换的酶不会导致氨基糖苷类药物的 MIC 值升高。

  DOI：

  10.1128/aac.01444-09

文献信息

• Simultaneous Quantification of Metabolites Involved in Central Carbon and Energy Metabolism Using Reversed-Phase Liquid Chromatography−Mass Spectrometry and in Vitro ¹³C Labeling
  作者：Wen-Chu Yang、Miroslav Sedlak、Fred E. Regnier、Nathan Mosier、Nancy Ho、Jiri Adamec

  DOI：10.1021/ac801693c

  日期：2008.12.15

  Comprehensive analysis of intracellular metabolites is a critical component of elucidating cellular processes. Although the resolution and flexibility of reversed-phase liquid chromatography−mass spectrometry (RPLC−MS) makes it one of the most powerful analytical tools for metabolite analysis, the structural diversity of even the simplest metabolome provides a formidable analytical challenge. Here we describe a robust RPLC−MS method for identification and quantification of a diverse group of metabolites ranging from sugars, phosphosugars, and carboxylic acids to phosphocarboxylics acids, nucleotides, and coenzymes. This method is based on in vitro derivatization with a 13C-labeled tag that allows internal standard based quantification and enables separation of structural isomer pairs like glucose 6-phosphate and fructose 6-phosphate in a single chromatographic run. Calibration curves for individual metabolites showed linearity ranging over more than 2 orders of magnitude with correlation coefficients of R2 > 0.9975. The detection limits at a signal-to-noise ratio of 3 were below 1.0 μM (20 pmol) for most compounds. Thirty common metabolites involved in glycolysis, the pentose phosphate pathway, and tricarboxylic acid cycle were identified and quantified from yeast lysate with a relative standard deviation of less than 10%.

  详细分析细胞内代谢物是阐明细胞过程的关键组成部分。尽管反相液相色谱-质谱联用（RPLC-MS）的分辨率和灵活性使其成为代谢物分析中最强大的分析工具之一，但即使是简单的代谢组，其结构的多样性也带来了巨大的分析挑战。本文描述了一种稳健的RPLC-MS方法，用于鉴定和定量一系列广泛的代谢物，包括糖类、磷酸糖、羧酸、磷酸羧酸、核苷酸和辅酶。该方法基于使用13C标记标签的体外衍生化，允许基于内标的定量，并能在单次色谱运行中分离结构异构体对，如葡萄糖6-磷酸和果糖6-磷酸。单个代谢物的校准曲线显示线性范围超过两个数量级，相关系数R² > 0.9975。大多数化合物的信噪比为3的检测限低于1.0 μM（20 pmol）。从酵母裂解液中鉴定和定量了涉及糖酵解、戊糖磷酸途径和三羧酸循环的30种常见代谢物，相对标准偏差小于10%。
• Chemical and enzymatic characterization of recombinant rabbit muscle pyruvate kinase
  作者：Christian Boehme、Frank Bieber、Julia Linnemann、Reinhard Breitling、Stefan Lorkowski、Siegmund Reissmann

  DOI：10.1515/hsz-2012-0334

  日期：2013.5.1

  synthesis of TTP via an enzymatic cascade reaction. The metalloenzyme PK shows pronounced promiscuity and therefore fits well to the conditions of this reaction. PK commonly used today is isolated from rabbit muscle. We cloned and expressed the respective open reading frame in Escherichia coli, purified, and characterized the His-tagged recombinant enzyme. The enzyme has an activity optimum at 37°C and

  摘要 胸苷三磷酸 (TTP) 的逐步合成在最后一步需要激酶进行磷酸化。因为使用磷酸烯醇式丙酮酸 (PEP) 作为底物的丙酮酸激酶 (PK) 可以再生三磷酸腺苷和磷酸化胸苷二磷酸，我们选择这种酶通过酶促级联反应合成 TTP。金属酶 PK 表现出明显的混杂性，因此非常适合该反应的条件。今天常用的PK是从兔肌肉中分离出来的。我们在大肠杆菌中克隆并表达了各自的开放阅读框，纯化并表征了带有 His 标签的重组酶。该酶在 37°C 和 7.4 至 7.8 的 pH 范围内具有最佳活性。Km 常数与分离的二磷酸腺苷 (ADP) 天然酶一致，为 0.37±0。02 毫米，PEP 为 0.07±0.01 毫米。重组酶在其底物特异性方面表现出以下范围：ADP>dADP>dGDP>dCDP>胸苷二磷酸(TDP)。它允许以高产率（高达 95%）将 TDP 磷酸化为 TTP。金属离子 Mg2+ 和 K+ 是完全酶活性所必需的。添加过渡金属离子如
• Prebiotic synthesis of phosphoenol pyruvate by α-phosphorylation-controlled triose glycolysis
  作者：Adam J. Coggins、Matthew W. Powner

  DOI：10.1038/nchem.2624

  日期：2017.4

  highest-energy phosphate found in living organisms and is one of the most versatile molecules in metabolism. Consequently, it is an essential intermediate in a wide variety of biochemical pathways, including carbon fixation, the shikimate pathway, substrate-level phosphorylation, gluconeogenesis and glycolysis. Triose glycolysis (generation of ATP from glyceraldehyde 3-phosphate via phosphoenol pyruvate)

  磷酸烯醇丙酮酸是在生物体中发现的能量最高的磷酸盐，并且是新陈代谢中用途最广泛的分子之一。因此，它是多种生化途径中必不可少的中间体，包括碳固定，the草酸酯途径，底物水平的磷酸化，糖异生和糖酵解。三糖糖酵解（通过磷酸烯醇丙酮酸由3-磷酸甘油醛生成ATP）是新陈代谢中最重要，最保守的途径之一。在这里，我们证明了从益生元核苷酸前体，乙醇醛和甘油醛高效有效地合成丙酮酸磷酸烯醇酯。此外，磷酸烯醇丙酮酸是在α-磷酸化控制的反应网络中衍生的，该网络可接触2-磷酸甘油酸，3-磷酸甘油酸，磷酸丝氨酸和丙酮酸。我们的结果表明，在温和的，益生元的合理条件下，能量转导和氨基酸，糖，核苷酸和脂质生物合成关键的核心代谢途径的关键成分可以高产量重建。
• Arabinose 5-phosphate analogues as mechanistic probes for Neisseria meningitidis 3-deoxy-d-manno-octulosonate 8-phosphate synthase
  作者：Meekyung Ahn、Fiona C. Cochrane、Mark L. Patchett、Emily J. Parker

  DOI：10.1016/j.bmc.2008.09.056

  日期：2008.11

  3-Deoxy-D-manno-octulosonate 8-phosphate (KDO8P) synthase catalyses the condensation reaction between phosphoenolpyruvate and D-arabinose 5-phosphate (D-A5P) in a key step in lipopolysaccharide biosynthesis in Gram-negative bacteria. The KDO8P synthase from Neisseria meningitidis was cloned into Escherichia coli, overexpressed and purified. A variety of D-A5P stereoisomers were tested as substrates

  3-脱氧-D-甘露糖八酸八磷酸酯（KDO8P）合酶催化磷酸烯醇丙酮酸和D-阿拉伯糖5-磷酸酯（D-A5P）之间的缩合反应，这是革兰氏阴性细菌中脂多糖生物合成的关键步骤。来自脑膜炎奈瑟氏球菌的KDO8P合酶被克隆到大肠杆菌中，过表达和纯化。测试了多种D-A5P立体异构体作为底物，其中只有D-A5P和1-X5P是底物。构建了脑膜炎奈瑟氏球菌KDO8P合酶的Asn59Ala突变体，该突变体保留了不到1％的野生型活性。这些结果与该酶的催化机理一致，其中D-A5P和Asn59的C2和C3羟基至关重要。
• Phosphoenolpyruvate carboxykinase from T. cruzi magnetic beads affinity-based screening assays on crude plant extracts from Brazilian Cerrado
  作者：Bruno S. do Amaral、Larissa R.G. da Silva、Alessandra L. Valverde、Lorena R.F. de Sousa、Richele P. Severino、Dulce H.F. de Souza、Quezia B. Cass

  DOI：10.1016/j.jpba.2020.113710

  日期：2021.1

  high-resolution mass spectrometry experiments. Senecic acid, syneilesinolide A, phytosphingosine and vanillic acid 4-glucopyranoside are herein reported for the first time for Q. grandiflora, D. burchellii, A. falcata, respectively. In addition, the specificity of the assay was observed since only catechin was fished out from the ethanolic extract of B. coccolobifolia leaves, despite the presence of

  在南美锥虫病的致病性克鲁维氏菌中，磷酸烯醇丙酮酸羧化激酶（TC PEPCK）与碳水化合物的分解代谢有关。由于其在寄生虫代谢中的重要性，它已成为开发抗南美锥虫病新药的有希望的目标。为了研究不同的配体筛选方法，将Tc PEPCK固定在胺封端的磁珠（Tc PEPCK-MB）上，并通过液相色谱串联质谱活性分析和K Mapp进行动力学表征以草酰乙酸为底物的10±1μM值。天然产物库通过其次生代谢产物提供了高度多样化的分子框架，在此描述了一种配体捕捞Tc PEPCK-MB分析法，用于在巴西Cerrado植物的四种乙醇提取物中寻找配体：Qualea grandiflora（Vochysiaceae），Diospyros burchellii（Ebenaceae），Anadenanthera falcata（豆科）和球孢白桦（大头藻科）。通过液相色谱串联高分辨率质谱实验对11种已鉴定的配体进行了化学表征。