2,3-Dihydroxybenzoyl 5'-adenylate(1-)

• 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: 2,3-Dihydroxybenzoyl 5'-adenylate(1-)
• 英文别名: [(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methyl (2,3-dihydroxybenzoyl) phosphate
• 化学式: C17H17N5O10P-
• 分子量: 482.3
• InChiKey: ULPVJDOMCRTJSN-RVXWVPLUSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  -1.5
• 重原子数：
  33
• 可旋转键数：
  7
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.29
• 拓扑面积：
  235
• 氢给体数：
  5
• 氢受体数：
  14

反应信息

• 作为反应物：
  描述：

  2,3-Dihydroxybenzoyl 5'-adenylate(1-) 、 L-seryl-AMP(1-) 生成 adenosine 5'-monophosphate 、 Enterobactin(1-) 、 氢(+1)阳离子
  参考文献：
  名称：

  摘要：

  DOI：
• 作为产物：
  描述：

  2,3-dihydroxybenzoate 、 adenosine 5'-triphosphate 、 氢(+1)阳离子 生成 2,3-Dihydroxybenzoyl 5'-adenylate(1-) 、 pyrophosphoric acid
  参考文献：
  名称：

  The EntF and EntE adenylation domains of Escherichia coli enterobactin synthetase: Sequestration and selectivity in acyl-AMP transfers to thiolation domain cosubstrates

  摘要：

  肠杆菌的三个蛋白质（EntE，B，F）六模块非核糖肽合成酶（NRPS）合成了三聚体（三个N-(2,3-二羟基苯甲酰)丝氨酸）三内酯类铁载体enterobactin。在这项工作中，142 kDa的四域蛋白EntF被分成两个双域片段：一个108 kDa的缩合和腺苷酸化构造EntF C-A，以及一个37 kDa的肽载体蛋白（PCP）和硫酯酶蛋白EntF PCP-TE。EntF C-A的腺苷酸化域活性形成丝氨酸-AMP，但失去了将丝氨酸基团转移给相应的EntF PCP-TE的能力。观察到将丝氨酸转移给异源PCP蛋白片段（表面活性素合成酶的SrfB1 PCP和yersiniabactin合成酶的Ybt PCP1）的速率分别为0.5 min^-1和0.01 min^-1。通过测量腺苷酸化域释放的焦磷酸二酯（PPi）的催化周转率来确定这些缓慢的酰化速率是否反映了酰/氨酰-AMP的解离，随后由异源PCP在溶液中的偶发硫酰化。完整的SrfB1 PCP蛋白质以及Ybt PCP1并没有刺激EntF C-A或EntE释放PPi的增加。因此，NRPS组装线的A和PCP域片段之间的氨酰化在转移是否真正发生在蛋白质域之间还是由于缓慢的氨酰-AMP释放和随后的非酶促硫酸盐捕获，必须经过动力学审查。

  DOI：

  10.1073/pnas.040572897

文献信息

• Kinetic and Inhibition Studies of Dihydroxybenzoate-AMP Ligase from <i>Escherichia coli</i>
  作者：Alison L. Sikora、Daniel J. Wilson、Courtney C. Aldrich、John S. Blanchard

  DOI：10.1021/bi100350c

  日期：2010.5.4

  biosynthetic pathways in pathogenic bacteria represents a promising strategy for antibacterial drug development. Escherichia coli synthesize and secrete the small molecule iron chelator siderophore, enterobactin, in response to intracellular iron depletion. Here we describe a detailed kinetic analysis of EntE, one of six enzymes in the enterobactin synthetase gene cluster. EntE catalyzes the ATP-dependent

  抑制病原菌中的铁载体生物合成途径是抗菌药物开发的一种有前景的策略。大肠杆菌合成并分泌小分子铁螯合铁载体肠杆菌素，以响应细胞内铁耗竭。在这里，我们描述了肠杆菌素合成酶基因簇中六种酶之一 EntE 的详细动力学分析。EntE 催化 2,3-二羟基苯甲酸 (DHB) 和磷酸泛酰巯基乙胺化 EntB (holo-EntB) 的 ATP 依赖性缩合形成共价芳基化 EntB，该产物对肠杆菌素的最终组装至关重要。初始速度研究表明，EntE 是通过 bi-uni-uni-bi 乒乓动力学机制进行的，k cat等于 2.8 s -1和DHB、ATP 和全息-EntB-ArCP 的K m值分别为 2.5、430 和 2.9 μM。抑制和直接结合实验表明，在第一个半反应（腺苷酸化）期间，DHB 首先与游离酶结合，然后是 ATP 和焦磷酸盐的释放，形成腺苷酸中间体。在第二个半反应（连接）期间，磷酸泛酰巯基乙胺化 EntB
• A protein interaction surface in nonribosomal peptide synthesis mapped by combinatorial mutagenesis and selection
  作者：Jonathan R. Lai、Michael A. Fischbach、David R. Liu、Christopher T. Walsh

  DOI：10.1073/pnas.0601038103

  日期：2006.4.4

  biosynthetic intermediates, displayed via covalent attachment to carrier proteins, by catalytic domains is critical for NRPS and polyketide synthase function. We report the use of combinatorial mutagenesis coupled with in vivo selection for the production of the Escherichia coli NRPS product enterobactin to map the surface of the aryl carrier protein (ArCP) domain of EntB that interacts with the downstream

  非核糖体肽合成酶（NRPS）和聚酮化合物合成酶是大型的多域酶，可以生物合成许多药学上重要的天然产物。催化结构域通过共价附着于载体蛋白上而表现出的生物合成中间体的识别对于NRPS和聚酮化合物合酶的功能至关重要。我们报告结合诱变结合体内选择用于生产大肠杆菌NRPS产品enteractactin来映射与下游延伸模块EntF相互作用的EntB的芳基载体蛋白（ArCP）结构域的表面。通过shot弹枪丙氨酸扫描生成了跨越EntB-ArCP的预测螺旋2和环2 /螺旋3的两个文库，并选择了它们支持肠杆菌素生产的能力。从尚存的水池中 我们鉴定了几个高度保守的疏水残基（M249，F264和A268）。在结构模型中，这些残基聚集在磷酸泛素化的丝氨酸附近，并且这些位置中的两个位于预测的螺旋3区域中。随后的体外研究与以下假设一致：这些残基在EntB上形成与EntF相互作用所需的表面。这些结果表明，螺旋3是EntB-
• Reconstitution and Characterization of the <i>Escherichia coli</i> Enterobactin Synthetase from EntB, EntE, and EntF
  作者：Amy M. Gehring、Ichiro Mori、Christopher T. Walsh

  DOI：10.1021/bi9726584

  日期：1998.2.1

  The siderophore molecule enterobactin, a cyclic trimeric lactone of N-(2,3-dihydroxybenzoyl)serine, is synthesized and secreted by Escherichia coli in response to iron starvation. Here we report the first reconstitution of enterobactin synthetase activity from pure protein components: holo-EntB, EntE, and holo-EntF. Holo-EntB and holo-EntF were obtained by pretreatment of apo-EntB and apo-EntF with

  响应铁饥饿，铁载体分子肠杆菌素（一种N-（2,3-二羟基苯甲酰基）丝氨酸的环状三聚内酯）被大肠杆菌合成和分泌。在这里，我们报道了从纯蛋白质成分：holo-EntB，EntE和holo-EntF首次重建肠杆菌素合成酶活性。Holo-EntB和holo-EntF是通过用辅酶A和EntD预处理apo-EntB和apo-EntF而获得的，从而消除了肠杆菌素合成酶中对EntD的需求。完整的EntF单体可充当催化剂，使用ATP，L-丝氨酸和酰基-完整的EntB在肠杆菌素中形成三个酰胺键和三个酯键，并由EntE用2,3-二羟基苯甲酸酯酰化，作为底物周转速度为120-140分钟-1。没有证据表明肠杆菌素合成酶组分具有稳定的复合物。推定的活性位点丝氨酸残基（Ser1138到Ala）的硫酯酶结构域中的Holo-EntF突变消除了肠杆菌素合成酶的活性。但是，突变体holo-EntF保留了丝氨酸与其连接的磷酸泛肽
• Enterobactin Biosynthesis in <i>Escherichia coli</i>:  Isochorismate Lyase (EntB) Is a Bifunctional Enzyme That Is Phosphopantetheinylated by EntD and Then Acylated by EntE Using ATP and 2,3-Dihydroxybenzoate
  作者：Amy M. Gehring、Kenneth A. Bradley、Christopher T. Walsh

  DOI：10.1021/bi970453p

  日期：1997.7.1

  In Escherichin coli, the siderophore molecule enterobactin is synthesized in response to iron deprivation by formation of an amide bond between 2,3-dihydroxybenzoate (2,3-DHB) and L-serine and formation of eater linkages between three such N-acylated serine residues, We show that EntB, previously described as the isochorismate lyase required for production of 2,3-DHB, is a bifunctional protein that also serves as an aryl carrier protein (ArCP) with a role in enterobactin assembly, EntB is phosphopantetheinylated near the C terminus in a reaction catalyzed by EntD with a k(cat) of 5 min(-1) and a K-m for apo-EntB of 6.5 mu M. This holo-EntB is then acylated with 2,3-DHB in a reaction catalyzed by EntE, previously described as the 2,3-DHB-AMP ligase, with a k(cat) of 100 min(-1) and a k(m) of <1 mu M for holo-EntB, The N-terminal 187 amino acids of EntB (isochorismate lyase domain) are not needed for reaction of EntB with either EntD or EntE as demonstrated by the equivalent catalytic efficiencies of the full-length EntB (residues 1-285) and the C-terminal EntB ArCP domain (residues 188-285) as substrates for both EntD and EntE.
• Vibriobactin Biosynthesis in <i>Vibrio cholerae</i>:  VibH Is an Amide Synthase Homologous to Nonribosomal Peptide Synthetase Condensation Domains
  作者：Thomas A. Keating、C. Gary Marshall、Christopher T. Walsh

  DOI：10.1021/bi001651a

  日期：2000.12.1

  The Vibrio cholerae siderophore vibriobactin is biosynthesized from three molecules of 2,3-dihydroxybenzoate (DHB), two molecules of L-threonine, and one of norspermidine. Of the four genes positively implicated in vibriobactin biosynthesis, we have here expressed, purified, and assayed the products of three: vibE, vibB, and vibH. All three are homologous to nonribosomal peptide synthetase (NRPS) domains: VibE is a 2,3-dihydroxybenzoate-adenosyl monophosphate Ligase, VibB is a bifunctional isochorismate lyase-aryl carrier protein (ArCP), and VibH is a novel amide synthase that represents a free-standing condensation (C) domain. VibE and VibB are homologous to EntE and EntB from Escherichia coli enterobactin synthetase; VibE activates DHB as the acyl adenylate and then transfers it to the free thiol of the phosphopantetheine arm of VibB's ArCP domain. VibH then condenses this DHB thioester (the donor) with the small molecule norspermidine (the acceptor), forming N-1-(2,3-dihydroxybenzoyl)norspermidine (DHB-NSPD) with a k(cat) of 600 min(-1) and a K-m for acyl-VibB of 0.88 muM and for norspermidine of 1.5 mM. Exclusive monoacylation of a primary amine of norspermidine was observed. VibH also tolerates DHB-acylated EntB and 1,7-diaminoheptane, octylamine, and hexylamine as substrates, albeit at lowered catalytic efficiencies. DHB-NSPD possesses one of three acylations required for mature vibriobactin, and its formation confirms VibH's role in vibriobactin biosynthesis. VibH is a unique NRPS condensation domain that acts upon an upstream carrier-protein-bound donor and a downstream amine, turning over a soluble amide product, in contrast to an archetypal NRPS-embedded C domain that condenses two carrier protein thioesters.