β-D-[6,6'-(3)H]fructose-2,6-bisphosphate | 1253393-43-9

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: β-D-[6,6'-(3)H]fructose-2,6-bisphosphate
• CAS: 1253393-43-9
• 化学式: C₆H₁₄O₁₂P₂
• 分子量: 344.102
• InChiKey: YXWOAJXNVLXPMU-JPCBLXIPSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -2.99
• 重原子数：
  20.0
• 可旋转键数：
  6.0
• 环数：
  1.0
• sp3杂化的碳原子比例：
  1.0
• 拓扑面积：
  203.44
• 氢给体数：
  7.0
• 氢受体数：
  8.0

反应信息

• 作为反应物：
  描述：

  β-D-[6,6'-(3)H]fructose-2,6-bisphosphate 在 乙二胺四乙酸 、 human hepatic 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, recombinant, with C-terminal hexa-His sequence 、 Cleland's reagent 、 bovine serum albumin 作用下， 反应 0.75h， 生成
  参考文献：
  名称：

  Enzymatic preparation of high-specific-activity β-d-[6,6′-3H]fructose-2,6-bisphosphate: Application to a sensitive assay for fructose-2,6-bisphosphatase

  摘要：

  beta-D-Fructose-2,6-bisphosphate (Fru-2,6-P-2) is an important regulator of eukaryotic glucose homeostasis, functioning as a potent activator of 6-phosphofructo-1-kinase and inhibitor of fructose-1,6-bisphosphatase. Pharmaceutical manipulation of intracellular Fru-2,6-P2 levels, therefore, is of interest for the treatment of certain diseases, including diabetes and cancer. [2-P-32]Fru-2,6-P2 has been the reagent of choice for studying the metabolism of this effector molecule; however, its short half-life necessitates frequent preparation. Here we describe a convenient, economical, one-pot enzymatic preparation of high-specific-activity tritium-labeled Fru-2,6-P2. The preparation involves conversion of readily available, carrier-free D-[6,6'-H-3]glucose to [6,6'-H-3]Fru-2,6-P-2 using hexokinase, glucose-6-phosphate isomerase, and 6-phosphofructo-2-kinase. The key reagent in this preparation, bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from human liver, was produced recombinantly in Escherichia coli and purified in a single step using an appendant C-terminal hexa-His affinity tag. Following purification by anion exchange chromatography using triethylammonium bicarbonate as eluant, radiochemically pure [6,6'-H-3]Fru-2,6-P-2 having a specific activity of 50 Ci/mmol was obtained in yields averaging 35%. [6,6'-H-3]Fru-2,6-P-2 serves as a stable, high-specific-activity substrate in a facile assay capable of detecting fructose-2,6-bisphosphatase in the range of 10(-14) to 10(-15) mol, and it should prove to be useful in many studies of the metabolism of this important biofactor. (C) 2010 Elsevier Inc. All rights reserved.

  DOI：

  10.1016/j.ab.2010.06.017
• 作为产物：
  描述：

  D-<6-³H>glucose 在 potassium dihydrogenphosphate 、 乙二胺四乙酸 、 yeast hexokinase 、 human hepatic 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, recombinant, with C-terminal hexa-His sequence 、 rabbit muscle glucose-6-phosphate isomerase 、 potassium chloride 、 2-(膦酰氧基)丙烯酸酯 、 5’-三磷酸腺苷 、 Cleland's reagent 、 bovine serum albumin 作用下， 反应 1.5h， 以40%的产率得到β-D-[6,6'-(3)H]fructose-2,6-bisphosphate
  参考文献：
  名称：

  Enzymatic preparation of high-specific-activity β-d-[6,6′-3H]fructose-2,6-bisphosphate: Application to a sensitive assay for fructose-2,6-bisphosphatase

  摘要：

  beta-D-Fructose-2,6-bisphosphate (Fru-2,6-P-2) is an important regulator of eukaryotic glucose homeostasis, functioning as a potent activator of 6-phosphofructo-1-kinase and inhibitor of fructose-1,6-bisphosphatase. Pharmaceutical manipulation of intracellular Fru-2,6-P2 levels, therefore, is of interest for the treatment of certain diseases, including diabetes and cancer. [2-P-32]Fru-2,6-P2 has been the reagent of choice for studying the metabolism of this effector molecule; however, its short half-life necessitates frequent preparation. Here we describe a convenient, economical, one-pot enzymatic preparation of high-specific-activity tritium-labeled Fru-2,6-P2. The preparation involves conversion of readily available, carrier-free D-[6,6'-H-3]glucose to [6,6'-H-3]Fru-2,6-P-2 using hexokinase, glucose-6-phosphate isomerase, and 6-phosphofructo-2-kinase. The key reagent in this preparation, bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from human liver, was produced recombinantly in Escherichia coli and purified in a single step using an appendant C-terminal hexa-His affinity tag. Following purification by anion exchange chromatography using triethylammonium bicarbonate as eluant, radiochemically pure [6,6'-H-3]Fru-2,6-P-2 having a specific activity of 50 Ci/mmol was obtained in yields averaging 35%. [6,6'-H-3]Fru-2,6-P-2 serves as a stable, high-specific-activity substrate in a facile assay capable of detecting fructose-2,6-bisphosphatase in the range of 10(-14) to 10(-15) mol, and it should prove to be useful in many studies of the metabolism of this important biofactor. (C) 2010 Elsevier Inc. All rights reserved.

  DOI：

  10.1016/j.ab.2010.06.017