cobalt(2+);3-[(1R,2S,3S,5Z,7S,8S,13S,15Z,17R,18R,19R)-2,7,18-tris(2-amino-2-oxoethyl)-3,13-bis(3-amino-3-oxopropyl)-17-[3-[[(2R)-2-hydroxypropyl]amino]-3-oxopropyl]-1,2,5,7,12,12,15,17-octamethyl-8,13,18,19-tetrahydro-3H-corrin-24-id-8-yl]propanamide

• 分子结构分类: 有机化合物 - 有机杂环化合物 - 四吡咯及其衍生物
• 英文名称: cobalt(2+);3-[(1R,2S,3S,5Z,7S,8S,13S,15Z,17R,18R,19R)-2,7,18-tris(2-amino-2-oxoethyl)-3,13-bis(3-amino-3-oxopropyl)-17-[3-[[(2R)-2-hydroxypropyl]amino]-3-oxopropyl]-1,2,5,7,12,12,15,17-octamethyl-8,13,18,19-tetrahydro-3H-corrin-24-id-8-yl]propanamide
• 化学式: C₄₈H₇₂N₁₁O₈*Co
• 分子量: 990.162
• InChiKey: GFVWZOGCSKVPRA-JFYQDRLCSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  2.52
• 重原子数：
  68
• 可旋转键数：
  20
• 环数：
  5.0
• sp3杂化的碳原子比例：
  0.67
• 拓扑面积：
  346
• 氢给体数：
  8
• 氢受体数：
  12

反应信息

• 作为反应物：
  描述：

  adenosine 5'-triphosphate 、 cobalt(2+);3-[(1R,2S,3S,5Z,7S,8S,13S,15Z,17R,18R,19R)-2,7,18-tris(2-amino-2-oxoethyl)-3,13-bis(3-amino-3-oxopropyl)-17-[3-[[(2R)-2-hydroxypropyl]amino]-3-oxopropyl]-1,2,5,7,12,12,15,17-octamethyl-8,13,18,19-tetrahydro-3H-corrin-24-id-8-yl]propanamide 、 10-[(2S,3S,4R)-5-[[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl]oxy-2,3,4-trihydroxypentyl]-7,8-dimethyl-5H-benzo[g]pteridine-2,4-diolate 生成 adenosylcob(III)inamide 、 氢(+1)阳离子 、 FAD 、 三磷酸酯
  参考文献：
  名称：

  鼠伤寒沙门氏菌的ATP：类卟啉腺苷转移酶的游离状态，与MgATP或与羟基钴胺素和MgATP的复合的三维结构。

  摘要：

  在鼠伤寒沙门氏菌中，腺苷钴胺素的生物合成途径中钴-碳键的形成是由cobA基因的产物催化的，该产物编码196个氨基酸残基的蛋白质。这种酶是一种ATP：co（I）类rinrinoid腺苷转移酶，可将腺苷部分从MgATP转移到广泛的co（I）rrinoid底物上，据信这些底物包括cobinamide，其前体cobyric酸以及可能尚未鉴定的其他物质，以及羟考巴兰。此处报道了CobA的三种X射线结构：无底物形式，CobA与MgATP的复合物以及CobA与MgATP和羟基钴胺素的三元复合物，分别达到2.1 A，1.8 A和2.1 A的分辨率。这些结构表明该酶是同源二聚体。在载脂蛋白结构中 多肽链从Arg（28）延伸到Lys（181），由从六链平行β-折叠（链顺序为324516）构建的α/β结构组成。此折叠的拓扑结构与RecA中观察到的非常相似蛋白质，解旋酶结构域，F（1）ATPase和腺苷酸科宾酰胺

  DOI：

  10.1021/bi002145o
• 作为产物：
  描述：

  diaquo-cobinamide 在 聚合甲醛 作用下， 反应 0.08h， 生成 cobalt(2+);3-[(1R,2S,3S,5Z,7S,8S,13S,15Z,17R,18R,19R)-2,7,18-tris(2-amino-2-oxoethyl)-3,13-bis(3-amino-3-oxopropyl)-17-[3-[[(2R)-2-hydroxypropyl]amino]-3-oxopropyl]-1,2,5,7,12,12,15,17-octamethyl-8,13,18,19-tetrahydro-3H-corrin-24-id-8-yl]propanamide
  参考文献：
  名称：

  Tetrazole derivatives and matrices as novel cobalamin coordinating compounds

  摘要：

  Cobalamin (Cbl, vitamin B-12) consists of two moieties: (i) the corrin ring with the central Co-ion in the oxidation states Co3+/2+/1+ and (ii) the nucleotide side chain. The lower position of the ring is typically occupied by the nucleotide base (Bzm), whereas the upper surface coordinates exchangeable ligands. We have found that amino-tetrazole can coordinate to H2O center dot Cbl (Co3+) with K-d = 10(-5)-10(-6) M. A specific group (presumably tetrazole, TZ) can be easily created in CNBr-activated Sepharose by treatment with N-3(-). The prepared matrix (STZ) contained approximate to 10 mM of the active groups, which bound H2O center dot corrinoids with K-d = 10(-5)-10(-6) M. Stability of STZ-Cbl bonds gradually increased and reached K-d = 10(-7) M over 10-20 h (20 degrees C, pH 6-7). This effect can be ascribed to partial displacement of Bzm and coordination of TZ to the lower position. The binding was most efficient at pH 4-7 and low ionic strength, yet, noticeable adsorption took place even at extreme conditions, pH 1-9 and I = 0-2 M. Reduced corrins (Co2+) also exhibited high affinity for STZ. The bound ligands could be eluted as H2O center dot Cbl (pH 0), HO center dot Cbl (pH 14) or diCN center dot Cbl (pH 9-12, CN-). The adsorbent is applicable for one-step purification of corrins from a crude extract; separation of aquo- and diaquo-forms; specific capturing of H2O center dot Cbl from a mixture containing organo-Cbls or protein-bound Cbl, analysis of peptide-Cbl dissociation kinetics, etc. (C) 2006 Elsevier B.V. All rights reserved.

  DOI：

  10.1016/j.jorganchem.2006.08.081

文献信息

• Purification and initial characterization of the ATP:corrinoid adenosyltransferase encoded by the cobA gene of Salmonella typhimurium
  作者：S Suh、J C Escalante-Semerena

  DOI：10.1128/jb.177.4.921-925.1995

  日期：1995.2

  The cobA gene of Salmonella typhimurium and its product were overexpressed to approximately 20% of the total cell protein. CobA was purified to 98% homogeneity; N-terminal sequence analysis (21 residues) of homogeneous protein confirmed the predicted amino acid sequence. ATP:corrinoid adenosyltransferase activity was demonstrated in vitro to be associated with CobA. This activity was optimal at pH 8 and 37 degrees C. A quantitative preference was determined for Mn(II) cations and ATP. The apparent Km of CobA for ATP was 2.8 microM, and that for cob(I)alamin was 5.2 microM. Vmax was measured at 0.43 nmol/min. Cobinamide served as the substrate for CobA to yield adenosylcobinamide. Activity was stable at 4 degrees C for several weeks but was lost rapidly at room temperature (50% overnight). Dithiothreitol was required to maintain the enzymatic activity of CobA.

  沙门氏菌的cobA基因及其产物被过度表达，占细胞总蛋白的约20％。CobA被纯化到98％的同质性；同质蛋白的N末端序列分析（21个氨基酸残基）确认了预测的氨基酸序列。体外实验证明，ATP：辅酶B12腺苷基转移酶活性与CobA相关。该活性在pH 8和37℃时最佳。 Mn（II）阳离子和ATP的定量偏好已确定。 CobA对ATP的表观Km为2.8微米，对cob（I）alamin的表观Km为5.2微米。 Vmax为0.43纳摩尔/分钟。 Cobinamide作为底物用于CobA，产生腺苷基cobinamide。该活性在4℃下稳定数周，但在室温下迅速丧失（过夜50％）。需要二硫苏糖醇来维持CobA的酶活性。
• Spectroscopic Characterization of Active-Site Variants of the PduO-type ATP:Corrinoid Adenosyltransferase from <i>Lactobacillus reuteri</i>: Insights into the Mechanism of Four-Coordinate Co(II)corrinoid Formation
  作者：Kiyoung Park、Paola E. Mera、Jorge C. Escalante-Semerena、Thomas C. Brunold

  DOI：10.1021/ic202096x

  日期：2012.4.16

  acid substitutions at seven residues that are involved in ATP-binding, an intersubunit salt bridge, and the hydrophobic region surrounding the bound corrin ring. A quantitative analysis of our MCD and EPR spectra indicates that the entire hydrophobic pocket below the corrin ring, and not just residue F112, is critical for the removal of the axial ligand from the cobalt center of the Co2+corrinoids. Our

  PduO 型 5'-三磷酸腺苷 (ATP)：来自罗伊氏乳杆菌( Lr PduO) 的类可啉腺苷转移酶催化 ATP 的腺苷基转移到 Co 1+钴胺素 (Cbl) 和 Co 1+ cobinamide (Cbi) 底物以分别合成腺苷钴胺素 (AdoCbl) 和腺苷钴胺 (AdoCbi + )。先前的研究表明，为了克服具有热力学挑战性的 Co 2+ → Co 1+还原，该酶会大大削弱轴向配体-Co 2+键，从而有效地生成四配位 (4c) Co 2+类可啉物质。探索如何LrPduO 产生这些不寻常的 4c 物种，我们使用了磁圆二色性 (MCD) 和电子顺磁共振 (EPR) 光谱技术。活性位点的氨基上形成图4c的Co的相对产率酸取代的效果2+类咕啉物种通过进行八个单氨基酸取代在中涉及的七个残基检查ATP结合，一个亚基间盐桥，并且围绕结合的 corrin 环的疏水区域。我们的 MCD 和 EPR 光谱的定量分析表明，corrin
• An in Vitro Reducing System for the Enzymic Conversion of Cobalamin to Adenosylcobalamin
  作者：Maris V. Fonseca、Jorge C. Escalante-Semerena

  DOI：10.1074/jbc.m102510200

  日期：2001.8

  Homogeneous ferredoxin (flavodoxin):NADP(+) reductase and flavodoxin A proteins served as electron donors for the reduction of co(III)rrinoids to co(I)rrinoids in vitro. The resulting co(I)rrinoids served as substrates for the ATP: co(I)rrinoid adenosyltransferase (CobA) enzyme of Salmonella enterica serovar Typhimurium LT2 and were converted to their respective adenosylated derivatives. The reaction products were isolated by reverse phase high performance liquid chromatography, and their identities were confirmed by UV-visible spectroscopy, mass spectrometry, and in vivo biological activity assays. Adenosylcobalamin generated by this system supported the activity of 1,2-propanediol dehydratase as effectively as authentic adenosylcobalamin. This is the first report of a protein system that can be coupled to the adenosyltransferase CobA enzyme for the conversion of co(III)rrinoids to their adenosylated derivatives.
• Vitols E.; Walker G.A.; Huennekens F.M., J Biol Chem, 1966, 0021-9258, 1455-61
  作者：Vitols E.、Walker G.A.、Huennekens F.M.

  DOI：——

  日期：——
• Purification and Initial Biochemical Characterization of ATP:Cob(I)alamin Adenosyltransferase (EutT) Enzyme of Salmonella enterica
  作者：Nicole R. Buan、Jorge C. Escalante-Semerena

  DOI：10.1074/jbc.m603069200

  日期：2006.6

  ATP:cob(I) alamin adenosyltransferase (EutT) of Salmonella enterica was overproduced and enriched to similar to 70% homogeneity, and its basic kinetic parameters were determined. Abundant amounts of EutT protein were produced, but all of it remained insoluble. Soluble active EutT protein ( similar to 70% homogeneous) was obtained after treatment with detergent. Under conditions in which cobalamin (Cbl) was saturating, K-m(ATP) = 10 mu M, k(cat) = 0.03 s(-1), and V-max = 54.5 nM min(-1). Similarly, under conditions in which MgATP was saturating, K-m(Cbl) = 4.1 mu M, k(cat) = 0.06 s(-1), and V-max = 105 nM min(-1). Unlike other ATP: co(I)rrinoid adenosyltransferases in the cell (i.e. CobA and PduO), EutT activity was >= 50-fold higher with ATP versus GTP, and EutT retained 80% of its activity with ADP substituted for ATP and was completely inactive with AMP as substrate, indicating that the enzyme requires the beta-phosphate group of the nucleotide substrate. The data suggest that the amino group of adenine might play a role in nucleotide recognition and/or binding. Unlike the housekeeping CobA enzyme, EutT was not inhibited by inorganic tripolyphosphate (PPPi). Results from P-31 NMR spectroscopy studies identified PPi and P-i as by-products of the EutT reaction. In the absence of Cbl, EutT cleaved ATP into adenosine and PPPi, suggesting that PPPi is broken down into PPi and P-i. Electron transfer protein partners for EutT were not encoded by the eut operon. EutT-dependent activity was detected in cell-free extracts of cobA strains enriched for EutT when FMN and NADH were used to reduce cob(III) alamin to cob(I) alamin.