(E)-3-Methylglutaconyl-1-CoA

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 脂肪酰基
• 英文名称: (E)-3-Methylglutaconyl-1-CoA
• 英文别名: (E)-5-[2-[3-[[(2R)-4-[[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-4-hydroxy-3-phosphonatooxyoxolan-2-yl]methoxy-oxidophosphoryl]oxy-oxidophosphoryl]oxy-2-hydroxy-3,3-dimethylbutanoyl]amino]propanoylamino]ethylsulfanyl]-3-methyl-5-oxopent-3-enoate
• 化学式: C27H37N7O19P3S-5
• 分子量: 888.6
• InChiKey: GXKSHRDAHFLWPN-RKYLSHMCSA-I

计算性质

• 辛醇/水分配系数(LogP)：
  -4.8
• 重原子数：
  57
• 可旋转键数：
  21
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.59
• 拓扑面积：
  440
• 氢给体数：
  5
• 氢受体数：
  24

反应信息

• 作为产物：
  描述：

  (S)-Hmg-CoA 生成 (E)-3-Methylglutaconyl-1-CoA 、 水
  参考文献：
  名称：

  Enoyl-CoA hydratase

  摘要：

  Enoyl-CoA hydratase (ECH) catalyzes the second step in the physiologically important beta-oxidation pathway of fatty acid metabolism. This enzyme facilitates the syn-addition of a water molecule across the double bond of a trans-2-enoyl-CoA thioester, resulting in the formation of a beta-hydroxyacyl-CoA thioester. The catalytic mechanism of this proficient enzyme has been studied in great depth through a combination of kinetic, spectroscopic, and structural techniques, and is proposed to occur via the formation of a single transition state. Sequence alignment and mutagenesis studies have implicated the key residues important for catalysis: Gly-141, Glu-144, and Glu-164 (rat liver ECH numbering). The two catalytic glutamic acid residues are believed to act in concert to activate a water molecule, while Gly-141 is proposed to be involved in substrate activation. Recently, two potent inhibitors of ECH have been reported in the literature, which result in the irreversible inactivation of the enzyme via covalent adduct formation. This review summarizes studies on the structure, mechanism, and inhibition of ECH. (C) 2002 Elsevier Science Ltd. All rights reserved.

  DOI：

  10.1016/s0968-0896(02)00333-4

文献信息

• Characterization of the Mycobacterial Acyl-CoA Carboxylase Holo Complexes Reveals Their Functional Expansion into Amino Acid Catabolism
  作者：Matthias T. Ehebauer、Michael Zimmermann、Arjen J. Jakobi、Elke E. Noens、Daniel Laubitz、Bogdan Cichocki、Hedia Marrakchi、Marie-Antoinette Lanéelle、Mamadou Daffé、Carsten Sachse、Andrzej Dziembowski、Uwe Sauer、Matthias Wilmanns

  DOI：10.1371/journal.ppat.1004623

  日期：——

  Biotin-mediated carboxylation of short-chain fatty acid coenzyme A esters is a key step in lipid biosynthesis that is carried out by multienzyme complexes to extend fatty acids by one methylene group. Pathogenic mycobacteria have an unusually high redundancy of carboxyltransferase genes and biotin carboxylase genes, creating multiple combinations of protein/protein complexes of unknown overall composition and functional readout. By combining pull-down assays with mass spectrometry, we identified nine binary protein/protein interactions and four validated holo acyl-coenzyme A carboxylase complexes. We investigated one of these - the AccD1-AccA1 complex from Mycobacterium tuberculosis with hitherto unknown physiological function. Using genetics, metabolomics and biochemistry we found that this complex is involved in branched amino-acid catabolism with methylcrotonyl coenzyme A as the substrate. We then determined its overall architecture by electron microscopy and found it to be a four-layered dodecameric arrangement that matches the overall dimensions of a distantly related methylcrotonyl coenzyme A holo complex. Our data argue in favor of distinct structural requirements for biotin-mediated γ-carboxylation of α−β unsaturated acid esters and will advance the categorization of acyl-coenzyme A carboxylase complexes. Knowledge about the underlying structural/functional relationships will be crucial to make the target category amenable for future biomedical applications.

  生物素介导的短链脂肪酸辅酶 A 酯羧化是脂质生物合成的一个关键步骤，由多酶复合物完成，使脂肪酸延伸一个亚甲基。致病分枝杆菌的羧基转移酶基因和生物素羧化酶基因具有异常高的冗余度，从而产生了多种蛋白质/蛋白质复合物组合，其总体组成和功能读数均不清楚。通过将牵引检测与质谱分析相结合，我们确定了九种二元蛋白质/蛋白质相互作用和四种有效的全酰基辅酶 A 羧化酶复合物。我们研究了其中一种--结核分枝杆菌中的 AccD1-AccA1 复合物，它的生理功能迄今未知。利用遗传学、代谢组学和生物化学方法，我们发现该复合体参与以甲基巴豆酰辅酶 A 为底物的支链氨基酸分解代谢。然后，我们通过电子显微镜确定了它的整体结构，发现它是一个四层十二分子排列，与远亲甲基巴豆酰辅酶 A 整体复合物的整体尺寸相吻合。我们的数据证明，生物素介导的α-β不饱和酸酯的γ-羧化需要不同的结构，这将推动酰辅酶A羧化酶复合物的分类。了解潜在的结构/功能关系对于将目标类别用于未来的生物医学应用至关重要。
• Purification and Characterization of 3-Methylcrotonyl-Coenzyme A Carboxylase from Higher Plant Mitochondria
  作者：C. Alban、P. Baldet、S. Axiotis、R. Douce

  DOI：10.1104/pp.102.3.957

  日期：1993.7.1

  3-Methylcrotonyl-coenzyme A (CoA) carboxylase was purified to homogeneity from pea (Pisum sativum L.) leaf and potato (Solanum tuberosum L.) tuber mitochondria. The native enzyme has an apparent molecular weight of 530,000 in pea leaf and 500,000 in potato tuber as measured by gel filtration. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate disclosed two nonidentical subunits. The larger subunit (B subunit) is biotinylated and has an apparent molecular weight of 76,000 in pea leaf and 74,000 in potato tuber. The smaller subunit (A subunit) is biotin free and has an apparent molecular weight of 54,000 in pea leaf and 53,000 in potato tuber. The biotin content of the enzyme is 1 mol/133,000 g of protein and 1 mol/128,000 g of protein in pea leaf and potato tuber, respectively. These values are consistent with an A4B4 tetrameric structure for the native enzyme. Maximal 3-methylcrotonyl-CoA carboxylase activity was found at pH 8 to 8.3 and at 35 to 38[deg]C in the presence of Mg2+. Kinetic constants (apparent Km values) for the enzyme substrates 3-methylcrotonyl-CoA, ATP, and HCO3- were: 0.1 mM, 0.1 mM, and 0.9 mM, respectively, for pea leaf 3-methylcrotonyl-CoA carboxylase and 0.1 mM, 0.07 mM, and 0.34 mM, respectively, for potato tuber 3-methylcrotonyl-CoA carboxylase. A steady-state kinetic analysis of the carboxylase-catalyzed carboxylation of 3-methylcrotonyl-CoA gave rise to parallel line patterns in double reciprocal plots of initial velocity with the substrate pairs 3-methylcrotonyl-CoA plus ATP and 3-methylcrotonyl-CoA plus HCO3- and an intersecting line pattern with the substrate pair HCO3- plus ATP. It was concluded that the kinetic mechanism involves a double displacement. Purified 3-methylcrotonyl-CoA carboxylase was inhibited by end products of the reaction catalyzed, namely ADP and orthophosphate, and by 3-hydroxy-3-methylglutaryl-CoA. Finally, as for the 3-methylcrotonyl-CoA carboxylases from mammalian and bacterial sources, plant 3-methylcrotonyl-CoA carboxylase was sensitive to sulfhydryl and arginyl reagents.

  CoA發現，在pH 8至8.3和35至38[deg]C的溫度下，在Mg2+的存在下，3-甲基巴豆酰辅酶A羧化酶的活性最高。 酶底物3-甲基巴豆酰辅酶A、ATP和HCO3-的動力學常數（表觀Km值）分別為：豌豆葉3-甲基巴豆酰辅酶A羧化酶為0.1 mM、0.1 mM和0.9 mM，馬鈴薯塊莖3-甲基巴豆酰辅酶A羧化酶為0.1 mM、0.07 mM和0.34 mM。 對羧化酶催化3-甲基巴豆酰辅酶A羧化的穩態動力學分析發現，在pH 8至8.3和35至38[deg]C的溫度下，在Mg2+的存在下，3-甲基巴豆酰辅酶A羧化酶的活性最高。 酶底物3-甲基巴豆酰辅酶A、ATP和HCO3-的動力學常數（表觀Km值
• Purification and Characterization of 3-Methylcrotonyl-Coenzyme-A Carboxylase from Leaves of Zea mays
  作者：T.A. Diez、E.S. Wurtele、B.J. Nikolau

  DOI：10.1006/abbi.1994.1141

  日期：1994.4

  purified to near homogeneity from maize leaves. The resulting preparations of 3-methylcrotonyl-CoA carboxylase have a specific activity of between 200 and 600 nmol.min-1.mg-1 protein, representing an approximately 5000-fold purification of the enzyme. The purified 3-methylcrotonyl-CoA carboxylase has a molecular weight of 853,000 +/- 34,000 and is composed of two types of subunits, a biotin-containing subunit

  3-甲基巴豆酰基辅酶A羧化酶已从玉米叶片中纯化至接近均一。所得的3-甲基巴豆酰基-CoA羧化酶制剂具有200至600nmol.min-1.mg-1蛋白的比活性，代表该酶的约5000倍纯化。纯化的3-甲基巴豆酰基-CoA羧化酶的分子量为853,000 +/- 34,000，由两种类型的亚基组成，一个含生物素的亚基为80 +/- 2 kDa，一个不含生物素的亚基为58.5 + /-1.5 kDa。这些数据表明该酶具有α6β6构型。活性的最佳pH为8.0。底物3-甲基巴豆酰基-CoA，ATP和HCO3-的动力学常数分别为11 microM，20 microM和0.8 mM。对具有可变浓度的两种底物的3-甲基巴豆酰基-CoA羧化酶反应的动力学研究证实，ATP和HCO3-依次与酶结合，并且ATP和3-甲基巴豆酰基-CoA以乒乓方式结合。但是，类似的分析表明，HCO3-在第一个位点的结合受到3-甲基巴豆酰基-CoA的影响。对Mg2
• Expression, purification, characterization of human 3-methylcrotonyl-CoA carboxylase (MCCC)
  作者：Ching-Hsuen Chu、Dong Cheng

  DOI：10.1016/j.pep.2007.01.012

  日期：2007.6

  The current study reports the use of baculovirus system to express functionally active human recombinant 3-methylcrotonyl-CoA carboxylase (MCCC), a heteromultimeric complex that is composed of alpha and beta subunits which are encoded by distinct genes. Using immuno-affinity purification, an efficient protocol has been developed to purify the active MCCC which appears to reside in a similar to 500-800 kDa complex in Superpose-6 gel-filtration chromatography. Consistent with the native enzyme, in the recombinant human MCCC, the stoichiometry of alpha and beta subunits are at a one:one ratio. The kc at value of the recombinant enzyme is determined to be similar to 4.0 s(-1). It also possesses Km values (ATP: 45 11 mu M; 3-methylcrotonyl-CoA: 74 7 mu M) similar to those reported for the native enzyme. The recombinant human MCCC described here may provide a counter-screen enzyme source for testing cross reactivity for inhibitors against acetyl-CoA carboxylases which are designed to treat obesity, type 2 diabetes and other metabolic disorders. (C) 2007 Elsevier Inc. All rights reserved.
• Biochemical characterization of human 3-methylglutaconyl-CoA hydratase and its role in leucine metabolism
  作者：Matthias Mack、Ute Schniegler-Mattox、Verena Peters、Georg F. Hoffmann、Michael Liesert、Wolfgang Buckel、Johannes Zschocke

  DOI：10.1111/j.1742-4658.2006.05218.x

  日期：——

  The metabolic disease 3‐methylglutaconic aciduria type I (MGA1) is characterized by an abnormal organic acid profile in which there is excessive urinary excretion of 3‐methylglutaconic acid, 3‐methylglutaric acid and 3‐hydroxyisovaleric acid. Affected individuals display variable clinical manifestations ranging from mildly delayed speech development to severe psychomotor retardation with neurological handicap. MGA1 is caused by reduced or absent 3‐methylglutaconyl‐coenzyme A (3‐MG‐CoA) hydratase activity within the leucine degradation pathway. The human AUH gene has been reported to encode for a bifunctional enzyme with both RNA‐binding and enoyl‐CoA‐hydratase activity. In addition, it was shown that mutations in the AUH gene are linked to MGA1. Here we present kinetic data of the purified gene product of AUH using different CoA‐substrates. The best substrates were (E)‐3‐MG‐CoA (Vmax = 3.9 U·mg−1, Km = 8.3 µm, kcat = 5.1 s−1) and (E)‐glutaconyl‐CoA (Vmax = 1.1 U·mg−1, Km = 2.4 µm, kcat = 1.4 s−1) giving strong evidence that the AUH gene encodes for the major human 3‐MG‐CoA hydratase in leucine degradation. Based on these results, a new assay for AUH activity in fibroblast homogenates was developed. The only missense mutation found in MGA1 phenotypes, c.719C>T, leading to the amino acid exchange A240V, produces an enzyme with only 9% of the wild‐type 3‐MG‐CoA hydratase activity.