[2-(13)C,2-(2)H]-O-deuteroglycolaldehyde | 1162168-83-3

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: [2-(13)C,2-(2)H]-O-deuteroglycolaldehyde
• CAS: 1162168-83-3
• 化学式: C₂H₄O₂
• 分子量: 63.0257
• InChiKey: WGCNASOHLSPBMP-ZSLVMXJJSA-N

反应信息

• 作为产物：
  描述：

  D-glyceraldehyde 3-phosphate 在 咪唑 、 wildtype triosephosphate isomerase from chicken muscle 、 重水 作用下， 生成 [2-(13)C]-O-deuteroglycolaldehyde 、 [2-(13)C,2-(2)H]-O-deuteroglycolaldehyde 、
  参考文献：
  名称：

  酶结构：磷酸丙糖异构酶催化的整个底物和片中底物的反应的转变状态非常相似

  摘要：

  已经为野生型和以下 TIM 突变体确定了 (R)-甘油醛 3-磷酸酯异构酶催化反应的 (kcat/Km)GAP 值和底物乙醇醛和 HPO32– 反应的 kcat/KHPiKGA 值： I172V、I172A、L232A 和 P168A（来自布氏锥虫的 TIM）；用于 208-YGGS 环 7 替换突变体的 208-TGAG（L7RM，来自鸡肌肉的 TIM）；和 Y208T、Y208S、Y208A、Y208F 和 S211A（酵母 TIM）。在野生型和大多数突变酶的动力学参数之间观察到极好的线性对数相关性，斜率为 1.04 ± 0.03，L232A 和 L7RM 具有正偏差。单位斜率表明，大多数突变导致整个底物和底物片催化反应的激活势垒发生相同的变化，从而通过与蛋白质催化剂的类似相互作用来稳定两个过渡态。这与二价阴离子作为活跃的旁观者的作用是一致的，它们将 TIM 保持在催化活性的笼状形式中。

  DOI：

  10.1021/ja501103b

文献信息

• Structural Mutations That Probe the Interactions between the Catalytic and Dianion Activation Sites of Triosephosphate Isomerase
  作者：Xiang Zhai、Tina L. Amyes、Rik K. Wierenga、J. Patrick Loria、John P. Richard

  DOI：10.1021/bi401019h

  日期：2013.8.27

  decreases in (kcat/Km)E and (kcat/Km)E·HPi, respectively, for deprotonation of GA catalyzed by free TIM and by the TIM·HPO32– complex. The mutation has little effect on the observed and intrinsic phosphite dianion binding energy or the magnitude of phosphite dianion activation of TIM for catalysis of deprotonation of GA. A loop 7 replacement mutant (L7RM) of TIM from chicken muscle was prepared by substitution

  磷酸丙糖异构酶 (TIM) 催化磷酸二羟基丙酮异构化形成d-甘油醛 3-磷酸。报道了 TIM 中的两个结构突变对催化截短底物乙醇醛 (GA) 反应的动力学参数和亚磷酸酯二价阴离子活化该反应的影响。P168A 突变导致 ( k cat / K m ) E和 ( k cat / K m ) E·HP i 的类似降低 50 倍和 80 倍，分别用于由游离 TIM 和 TIM·催化的 GA 去质子化HPO 3 2–复杂的。该突变对观察到的和固有的亚磷酸酯二价阴离子结合能或用于催化 GA 去质子化的 TIM 的亚磷酸酯二价阴离子活化程度几乎没有影响。通过用 208-YGGS 替换古菌序列 208-TGAG 制备了来自鸡肌肉的 TIM 环 7 置换突变体 (L7RM)。L7RM 的 ( k cat / K m ) E降低了 25 倍，( k cat / K m ) E·HP i降低了 170 倍GA
• Role of Lys-12 in Catalysis by Triosephosphate Isomerase: A Two-Part Substrate Approach
  作者：Maybelle K. Go、Astrid Koudelka、Tina L. Amyes、John P. Richard

  DOI：10.1021/bi100538b

  日期：2010.6.29

  We report that the K12G mutation in triosephosphate isomerase (TIM) from Saccharomyces cerevisiae results in (1) a similar to 50-fold increase in K-m for the substrate glyceraldehyde 3-phosphate (GAP) and a 60-fold increase in K-i for competitive inhibition by the intermediate analogue 2-phosphoglycolate, resulting from the loss of stabilizing ground state interactions between the alkylammonium side chain of Lys-12 and the ligand phosphodianion group; (2) a 12000-fold decrease in k(cat) for isomerization of GAP, suggesting a tightening of interactions between the side chain of Lys-12 and the substrate on proceeding from the Michaelis complex to the transition state; and (3) a 6 x 10(5)-fold decrease in k(cat)/K-m, corresponding to a total 7.8 kcal/mol stabilization of the transition state by the cationic side chain of Lys-12. The yields of the four products of the K12G TIM-catalyzed isomerization of GAP in D2O were quantified as dihydroxyacetone phosphate (D H A P) (27%), [1(R)-H-2]DHAP (23%), [2(R)-H-2]GAP (31%), and methylglyoxal (18%) from an enzyme-catalyzed elimination reaction. The K12G mutation has only a small effect on the relative yields of the three products of the transfer of a proton to the TIM-bound enediol(ate) intermediate in D2O, but it strongly favors catalysis of the elimination reaction to give methylglyoxal. The K12G mutation also results in a >= 14-fold decrease in k(cat)/K-m for isomerization of bound glycolaldehyde (GA), although the dominant observed product of the mutant enzyme-catalyzed reaction of [1-C-13]GA in D2O is [1-C-13,2,2-di-H-2]GA from a nonspecific protein-catalyzed reaction. The observation that the K12G mutation results in a large decrease in k(cat)/K-m for the reactions of both GAP and the neutral truncated substrate [1-C-13]GA provides evidence for a stabilizing interaction between the cationic side chain of Lys-12 and the negative charge that develops at the enolate-like oxygen in the transition state for deprotonation of the sugar substrate "piece".