4-Fumarylacetoacetate(2-)

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 酮酸及其衍生物
• 英文名称: 4-Fumarylacetoacetate(2-)
• 英文别名: (E)-4,6-dioxooct-2-enedioate
• 化学式: C8H6O6-2
• 分子量: 198.13
• InChiKey: GACSIVHAIFQKTC-OWOJBTEDSA-L

计算性质

• 辛醇/水分配系数(LogP)：
  0.6
• 重原子数：
  14
• 可旋转键数：
  4
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.25
• 拓扑面积：
  114
• 氢给体数：
  0
• 氢受体数：
  6

反应信息

• 作为反应物：
  描述：

  4-Fumarylacetoacetate(2-) 、 水 生成 acetoacetic acid anion 、 fumarate 、 氢(+1)阳离子
  参考文献：
  名称：

  碳-碳键水解酶的晶体结构和机理。

  摘要：

  背景技术富马酸乙酰乙酸酯水解酶（FAH）催化酪氨酸和苯丙氨酸分解代谢的最后步骤，富马酸乙酰乙酸酯中碳-碳键的水解裂解，产生富马酸酯和乙酰乙酸酯。FAH没有已知的序列同源物，其机制未知。碳-碳水解反应对于人类芳香族氨基酸的代谢至关重要。FAH缺乏会导致致命的遗传性疾病I型酪氨酸血症。作为全球碳循环的一部分，碳-碳键水解在芳香族化合物的微生物代谢中也很重要。结果FAH晶体结构已通过对多波长异常衍射数据的快速，自动分析来确定。FAH多肽折叠成120个残基的N端结构域和300个残基的C端结构域。C端结构域定义了不寻常的β-链拓扑结构和新颖的“混合β-三明治夹层卷”结构。还确定了FAH及其生理产物的结构。该结构揭示了富马酸酯结合到活性位点的入口附近，而乙酰乙酸酯结合到紧邻Glu-His dyad的八面体配位的钙离子。结论FAH代表了水解酶的第一个结构，该结构特别作用于碳-碳键。FAH还定义了一类新型的金属酶，其特征是独特的alpha

  DOI：

  10.1016/s0969-2126(99)80170-1
• 作为产物：
  描述：

  4-Maleylacetoacetate 生成 4-Fumarylacetoacetate(2-)
  参考文献：
  名称：

  Discovery of a functional polymorphism in human glutathione transferase zeta by expressed sequence tag database analysis

  摘要：

  通过序列比对对表达序列标签（EST）数据库进行分析，可以快速筛选出具有生理意义的蛋白质的多态性。最近对人类 zeta 类谷胱甘肽转移酶 GSTZ1 进行了鉴定，对表达序列标签克隆的分析表明该基因可能存在多态性。本报告确定了三种 GSTZ1 等位基因，它们是由编码区第 94 和 124 位核苷酸上的 A 到 G 转变引起的，即 GSTZ1*A - A94A124;GSTZ1*B - A94G124;GSTZ1*C - G94G124。对高加索对照人群（n = 141）进行的聚合酶链反应/限制性片段长度多态性分析表明，这三种等位基因都存在，Z1*A、Z1*B 和 Z1*C 的频率分别为 0.09、0.28 和 0.63。这些核苷酸替换均为非同义替换，其中第 94 和 124 位的 A 到 G 分别表示 Lys32 到 Glu 和 Arg42 到 Gly 的替换。变体蛋白在大肠杆菌中表达为 6X His 标记蛋白，并通过 Ni-agarose 柱色谱法纯化。对重组蛋白活性的研究发现，与 GSTZ1b-1b 和 GSTZ1c-1c 相比，GSTZ1a-1a 对几种底物的活性存在差异，其中对二氯乙酸的活性高出 3.6 倍。该报告表明，通过分析 EST 数据库，发现了一种功能性多态性。

  DOI：

  10.1097/00008571-200002000-00007

文献信息

• Mechanistic Inferences from the Crystal Structure of Fumarylacetoacetate Hydrolase with a Bound Phosphorus-based Inhibitor
  作者：Raynard L. Bateman、P. Bhanumoorthy、John F. Witte、Ronald W. McClard、Markus Grompe、David E. Timm

  DOI：10.1074/jbc.m007621200

  日期：2001.1

  human metabolic function, with loss of FAH activity causing the fatal metabolic disease hereditary tyrosinemia type I (HT1). An enzymatic mechanism involving a catalytic metal ion, a Glu/His catalytic dyad, and a charged oxyanion hole was previously proposed based on recently determined FAH crystal structures. Here we report the development and characterization of an FAH inhibitor, 4-(hydroxymethylph

  富马酸乙酰乙酸酯水解酶（FAH）催化富马酸乙酰乙酸酯中碳-碳键的水解裂解，生成富马酸酯和乙酰乙酸酯，作为Phe和Tyr降解的最后一步。这种不寻常的反应是人体的一项基本代谢功能，FAH活性下降会导致致命的代谢性疾病I型遗传性酪氨酸血症（HT1）。基于最近确定的FAH晶体结构，先前提出了涉及催化金属离子，Glu / His催化二元体和带电荷的氧阴离子孔的酶促机理。在这里，我们报告发展和表征的FAH抑制剂4-（羟甲基膦酰基）-3-氧代丁酸（HMPOBA），与生理底物竞争与K（i）为85 microM。与HMPOBA络合的FAH的晶体结构在1。3-A分辨率揭示了竞争抑制的分子基础，支持在FAH催化反应期间提议的四面体烷氧基过渡态中间体的形成，并揭示了在酶的活性位点结合的Mg（2+）。对应于不同催化状态的FAH结构的分析揭示了显着的活性位点侧链运动，这也可能与催化功能有关。因此，这些结果促进了对与致
• Characterization of the human fumarylacetoacetate hydrolase gene and identification of a missense mutation abolishing enzymatic activity
  作者：Yves Labelle、Daniel Phaneuf、Barbara Leclerc、Robert M.Tanguay

  DOI：10.1093/hmg/2.7.941

  日期：——

  Hereditary tyrosinemia type 1 is an autosomal recessive disease caused by a deficiency of the last enzyme in the catabollc pathway of tyrosine, fumarylacetoacetate hydrolase (FAH). To analyze the mutations involved in this disease, and as a first step towards elucidating the mechanisms regulating the transcription of the FAH gene, we have isolated and characterized the human gene coding for FAH. The gene contains 14 exons and spans approximately 35 kilobases of DNA. The 5' end of the gene is highly GC-rich, and eleven putative binding sites for the transcription factor Sp 1 were Identified In the proximal region of the promoter. We Investigated the molecular basis of FAH deficiency in a hereditary tyrosinemia type 1 patient whose liver FAH showed a very low enzymatic activity. Sequencing of the liver FAH cDNA of the patient revealed a C to A transversion in the FAH mRNA, which predicted the replacement of an alanine (A) residue with an aspartic acid (D) residue at position 134 (A134D) of the amino acid sequence of the corresponding protein. Direct sequencing of genomic DNA Indicated that the patient was heterozygous for the A134D mutation. The allele that does not carry the A134D mutation was expressed at a very low level in the liver of the patient. Expression of the mutant allele in CV-1 cells confirmed that the A134D mutation was responsible for the lack of enzymatic activity in the liver of the patient.

  遗传性酪氨酸血症 1 型是一种常染色体隐性遗传病，由酪氨酸代谢途径中的最后一种酶富马酸乙酰乙酸水解酶（FAH）缺乏引起。为了分析这种疾病所涉及的突变，并作为阐明 FAH 基因转录调控机制的第一步，我们分离并鉴定了编码 FAH 的人类基因。该基因包含 14 个外显子，横跨约 35 千碱基的 DNA。该基因的 5' 端高度富含 GC，在启动子的近端区域发现了 11 个转录因子 Sp 1 的推定结合位点。我们对一名肝脏 FAH 酶活性极低的遗传性酪氨酸血症 1 型患者的 FAH 缺乏症的分子基础进行了研究。对该患者肝脏 FAH cDNA 的测序发现，FAH mRNA 中存在一个 C 到 A 的反转，这预示着在相应蛋白质氨基酸序列的第 134 位（A134D），丙氨酸（A）残基被天冬氨酸（D）残基取代。基因组 DNA 的直接测序表明，患者是 A134D 突变的杂合子。不携带 A134D 突变的等位基因在患者肝脏中的表达量非常低。突变等位基因在 CV-1 细胞中的表达证实，A134D 突变是导致患者肝脏中缺乏酶活性的原因。
• Point mutations in the murine fumarylacetoacetate hydrolase gene: Animal models for the human genetic disorder hereditary tyrosinemia type 1
  作者：Jennifer L. Aponte、Gary A. Sega、Loren J. Hauser、Madhu S. Dhar、Catherine M. Withrow、Donald A. Carpenter、Eugene M. Rinchik、Cymbeline T. Culiat、Dabney K. Johnson

  DOI：10.1073/pnas.98.2.641

  日期：2001.1.16

  Hereditary tyrosinemia type 1 (HT1) is a severe autosomal recessive metabolic disease associated with point mutations in the human fumarylacetoacetate hydrolase ( FAH ) gene that disrupt tyrosine catabolism. An acute form of HT1 results in death during the first months of life because of hepatic failure, whereas a chronic form leads to gradual development of liver disease often accompanied by renal dysfunction, childhood rickets, neurological crisis, and hepatocellular carcinoma. Mice homozygous for certain chromosome 7 deletions of the albino Tyr ; c locus that also include Fah die perinatally as a result of liver dysfunction and exhibit a complex syndrome characterized by structural abnormalities and alterations in gene expression in the liver and kidney. Here we report that two independent, postnatally lethal mutations induced by N -ethyl- N -nitrosourea and mapped near Tyr are alleles of Fah . The Fah ^6287SB allele is a missense mutation in exon 6, and Fah ^5961SB is a splice mutation causing loss of exon 7, a subsequent frameshift in the resulting mRNA, and a severe reduction of Fah mRNA levels. Increased levels of the diagnostic metabolite succinylacetone in the urine of the Fah ^6287SB and Fah ^5961SB mutants indicate that these mutations cause a decrease in Fah enzymatic activity. Thus, the neonatal phenotype present in both mutants is due to a deficiency in Fah caused by a point mutation, and we propose Fah ^5961SB and Fah ^6287SB as mouse models for acute and chronic forms of human HT1, respectively.

  遗传性酪氨酸血症1型（HT1）是一种严重的常染色体隐性代谢性疾病，与人类烟酰乙酰乙酸水解酶（FAH）基因的点突变有关，破坏了酪氨酸的分解代谢。急性HT1导致肝功能衰竭，导致婴儿在出生后的前几个月死亡，而慢性HT1则导致肝脏疾病的逐渐发展，通常伴有肾功能障碍、儿童佝偻病、神经危机和肝细胞癌。小鼠染色体7缺失的白化病Tyr;c基因座中的某些纯合子，也包括FAh，因肝功能障碍而死亡，并表现出复杂的综合征，其特征为肝脏和肾脏的结构异常和基因表达的改变。在这里，我们报告了由N-乙基-N-亚硝基脲诱导的两个独立的产后致死突变，并在Tyr附近映射的突变体是FAh的等位基因。FAh 6287SB等位基因是外显子6的错义突变，而FAh 5961SB是一个剪接突变，导致外显子7的丧失，随后在所得到的mRNA中出现移码，并且FAh mRNA水平严重降低。在FAh 6287SB和FAh 5961SB突变体的尿液中增加的诊断代谢物丙酮琥珀酸酯表明，这些突变导致FAh酶活性降低。因此，两个突变体中出现的新生儿表型是由于点突变引起的FAh缺乏，我们建议FAh 5961SB和FAh 6287SB作为人类HT1急性和慢性形式的小鼠模型。
• A preliminary characterization of the cytosolic glutathione transferase proteome from <i>Drosophila melanogaster</i>
  作者：Chonticha Saisawang、Jantana Wongsantichon、Albert J. Ketterman

  DOI：10.1042/bj20111747

  日期：2012.2.15

  The cytosolic GST (glutathione transferase) superfamily has been annotated in the Drosophila melanogaster genome database. Of 36 genes, four undergo alternative splicing to yield a total of 41 GST proteins. In the present study, we have obtained the 41 transcripts encoding proteins by RT (reverse transcription)–PCR using RNA template from Drosophila S2 cells, an embryonic cell line. This observation suggests that all of the annotated DmGSTs (D. melanogaster GSTs) in the proteome are expressed in the late embryonic stages of D. melanogaster. To avoid confusion in naming these numerous DmGSTs, we have designated them following the universal GST nomenclature as well as previous designations that fit within this classification. Furthermore, in the cell line, we identified an apparent processed pseudogene, gste8, in addition to two isoforms from the Delta class that have been published previously. Only approximately one-third of the expressed DmGSTs could be purified by conventional GSH affinity chromatography. The diverse kinetic properties as well as physiological substrate specificity of the DmGSTs are such that each individual enzyme displayed a unique character even compared with members from the same class.

  黑腹果蝇基因组数据库已注释了细胞质 GST（谷胱甘肽转移酶）超家族。在 36 个基因中，有 4 个经过了替代剪接，总共产生了 41 个 GST 蛋白。在本研究中，我们利用果蝇 S2 细胞（一种胚胎细胞系）的 RNA 模板，通过 RT（反转录）-PCR 获得了这 41 个编码蛋白质的转录本。这一观察结果表明，蛋白质组中所有注释的 DmGSTs（黑腹果蝇 GSTs）都在黑腹果蝇的胚胎晚期表达。为了避免在命名这些数量众多的 DmGST 时出现混淆，我们按照通用的 GST 命名法以及以前符合这一分类的命名法对它们进行了命名。此外，在细胞系中，除了以前发表过的两种德尔塔类同工酶外，我们还发现了一种明显经过加工的假基因--gSTe8。只有大约三分之一表达的 DmGST 能通过传统的 GSH 亲和层析法纯化。DmGSTs 的动力学特性和生理底物特异性各不相同，因此，即使与同类成员相比，每个酶也显示出独特的特性。
• Glutathione transferase Zeta catalyses the oxygenation of the carcinogen dichloroacetic acid to glyoxylic acid
  作者：Zeen TONG、Philip G. BOARD、M. W. ANDERS

  DOI：10.1042/bj3310371

  日期：1998.4.15

  Dichloroacetic acid (DCA), a common drinking-water contaminant, is hepatocarcinogenic in rats and mice, and is a therapeutic agent used clinically in the management of lactic acidosis. DCA is biotransformed to glyoxylic acid by glutathione-dependent cytosolic enzymes in vitro and is metabolized to glyoxylic acid in vivo. The enzymes that catalyse the oxygenation of DCA to glyoxylic acid have not, however, been identified or characterized. In the present investigation, an enzyme that catalyses the glutathione-dependent oxygenation of DCA was purified to homogeneity (587-fold) from rat liver cytosol. SDS/PAGE and HPLC gel-filtration chromatography showed that the purified enzyme had a molecular mass of 27–28 kDa. Sequence analysis showed that the N-terminus of the purified protein was blocked. An internal sequence of 30 amino acid residues was obtained that matched the recently discovered human glutathione transferase Zeta well [Board, Baker, Chelvanayagam and Jermiin (1997) Biochem. J. 328, 929–935]. Western-blot analysis showed that the purified rat-liver enzyme cross-reacted with rabbit antiserum raised against recombinant human glutathione transferase Zeta. The apparent Km and Vmax values of the purified enzyme with DCA as the variable substrate were 71.4 µM and 1334 nmol/min per mg of protein, respectively; the Km for glutathione was 59 µM. Both the purified rat-liver enzyme and the recombinant human enzyme showed high activity with DCA as the substrate. These results demonstrate that the glutathione-dependent oxygenation of DCA to glyoxylic acid is catalysed by a Zeta-class glutathione transferase.

  二氯乙酸（DCA）是一种常见的饮用水污染物，在大鼠和小鼠中具有肝癌致癌作用，并且在临床上用于乳酸中毒的治疗。DCA在体外通过谷胱甘肽依赖性胞浆酶生物转化为乙醛酸，并在体内代谢为乙醛酸。然而，催化DCA氧化为乙醛酸的酶尚未被确定或表征。在本研究中，从大鼠肝细胞浆中纯化了一种催化谷胱甘肽依赖性氧化DCA的酶，纯度为587倍。SDS / PAGE和HPLC凝胶过滤色谱表明，纯化的酶的分子量为27-28 kDa。序列分析表明，纯化蛋白的N-末端被阻断。获得了一个30个氨基酸残基的内部序列，与最近发现的人类谷胱甘肽转移酶ζ（GSTZ）非常匹配。WeSTern-blot分析表明，纯化的大鼠肝酶与兔抗人重组谷胱甘肽转移酶ζ的血清交叉反应。纯化酶以DCA为可变底物的表观Km和Vmax值分别为71.4 µM和1334 nmol / min每毫克蛋白质，谷胱甘肽的Km为59 µM。纯化的大鼠肝酶和重组人酶均显示出高活性，以DCA为底物。这些结果表明，谷胱甘肽依赖性氧化DCA为乙醛酸是由ζ级谷胱甘肽转移酶催化的。