3'-<<2-(acetylamino)-2-deoxy-β-D-glucopyranosyl>oxy>-6'-hydroxyspiroxanthen>-3-one

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: 3'-<<2-(acetylamino)-2-deoxy-β-D-glucopyranosyl>oxy>-6'-hydroxyspiroxanthen>-3-one
• 英文别名: 3'-[[2-(acetylamino)-2-deoxy-β-D-glucopyranosyl]oxy]-6'-hydroxyspiro[isobenzofuran-1(3H),9'-[9H]-xanthen]-3-one；fluorescein mono-beta-D-(2-deoxy-2-N-acetyl)glucopyranoside；fluorescein mono-β-D-N-acetylglucosamine；FL-GIcNAc；FM-GlcNAc；3'-{[2-(acetylamino)-2-deoxy-β-D-glucopyranosyl]oxy}-6'-hydroxyspiro(isobenzofuran-1(3H),9'-[9H]xanthen)-3-one；Fluorescein mono-beta-D-N-acetylglucosamine；N-[(2S,3R,4R,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-(6'-hydroxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl)oxyoxan-3-yl]acetamide
• 化学式: C₂₈H₂₅NO₁₀
• 分子量: 535.507
• InChiKey: YHERABVROCVWLO-CIOJGTTESA-N

计算性质

• 辛醇/水分配系数(LogP)：
  1.6
• 重原子数：
  39
• 可旋转键数：
  4
• 环数：
  6.0
• sp3杂化的碳原子比例：
  0.29
• 拓扑面积：
  164
• 氢给体数：
  5
• 氢受体数：
  10

反应信息

• 作为产物：
  描述：

  荧光素 在 吡啶 、 ammonium hydroxide 、 silver(l) oxide 作用下， 以 甲醇 、 乙腈 为溶剂， 反应 51.0h， 生成 3'-<<2-(acetylamino)-2-deoxy-β-D-glucopyranosyl>oxy>-6'-hydroxyspiroxanthen>-3-one
  参考文献：
  名称：

  Enzymatic characterization of O-GlcNAcase isoforms using a fluorogenic GlcNAc substrate

  摘要：

  A highly sensitive fluorogenic hexosaminidase substrate, fluorescein di(N-acetyl-beta-D-glucosaminide) (FDGlcNAc) was prepared essentially as described previously [Chem. Pharm. Bull. 1993, 41, 314] with some modifications. The fluorescent analog is a substrate for a number of hexosaminidases but here we have focused on the cytoplasmic O-GlcNAcase isoforms. Kinetic analysis using purified O-GlcNAcase and its splice variant (v-O-GlcNAcase) expressed in Escherichia coli suggests that FDGlcNAc is a much more efficient substrate (K-m = 84.9 mu M) than the conventional substrate, para-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside (pNP-beta-GlcNAc, K-m = 1.1 nM) and a previously developed fluorogenic substrate, 4-methylumbelliferyl 2-acetamido-2-deoxy-beta -D-glucopyranoside [MUGlcNAc, K-m = 0.43 mM; J. Biol. Chem. 2005, 280, 25313] for O-GlcNAcase. The variant O-GlcNAcase. a protein lacking the C-terminal third of the full-length O-GlcNAcase, exhibited a K-m of 2.1 mM with respect to FDGlcNAc. This shorter isoforni was not previously thought to exhibit O-GlcNAcase activity based on in vitro Studies with pNP-beta-GlcNAc. However, both O-GlcNAcase isoforms reduced O-GlcNAc protein levels extracted from HeLa and HT-29 cells in vitro. indicating that the splice variant is a bona fide O-GlcNAcase. Fluorescein di-N-acetyl-beta-D-galactosaminide (FDGalNAc) is not cleaved by these enzymes, consistent with previous findings that the O-GlcNAcase has substrate specificity toward O-GlcNAc but not O-GalNAc. The enzymatic activity of the shorter isoforni of O-GlcNAcase was first detected by using highly sensitive fluorogenic FDGlcNAc substrate. The finding that O-GlcNAcase exists as two distinct isoforms has a number of important implications for the role of O-GlcNAcase in hexosamine signaling. Published by Elsevier Ltd.

  DOI：

  10.1016/j.carres.2006.03.004

文献信息

• Glycosides Having Chromophores as Substrates for Sensitive Enzyme Analysis. IV. Synthesis of N-Acetyl-.BETA.-D-glucosaminides of Fluorescein Derivatives and Their Application to the Rate-Assay of N-Acetyl-.BETA.-D-glucosaminidase.
  作者：Kouichi KASAI、Riichiro UCHIDA、Nobuyuki YAMAJI

  DOI：10.1248/cpb.41.314

  日期：——

  Three novel N-acetyl-β-D-glucosaminides, 2', 7'-dichlorofluorescein mono(N-acetyl-β-D-glucosaminide) (6a), fluorescein mono(N-acetyl-β-D-glucosaminide) (6b) and 2', 7'-dichlorofluorescein di(N-acetyl-β-D-glucosaminide) (7a), were synthesized by the introduction of N-acetyl-β-D-glucosaminyl group into fluorescein derivatives followed by the removal of the protecting group. Compounds 6a, 6b and 7a were hydrolyzed by N-acetyl-β-D-glucosaminidase to give products showing high absorbance at a long absorption wavelength (500, 465 and 485 nm) under weakly acidic rate-assay conditions (pH 5.0). The Km values for 6a and 7a were 0.56 and 0.86 mM, respectively. Among these compounds, 7a is considered to be the most potential chromogenic substrate for the rate-assay of N-acetyl-β-D-glucosaminidase, since it gives a clear color generation from colorless to orange color (λmax 280→485 nm) by enzyme hydrolysis and has a higher water solubility of more than 30 mM.

  通过引入 N-乙酰基-β-D-氨基葡萄糖苷，合成了三种新型 N-乙酰基-β-D-氨基葡萄糖苷，即 2'，7'-二氯荧光素单(N-乙酰基-β-D-氨基葡萄糖苷)(6a)、荧光素单(N-乙酰基-β-D-氨基葡萄糖苷)(6b)和 2'、7'-dichlorofluorescein di(N-acetyl-β-D-glucosaminide) (7a)是通过在荧光素衍生物中引入 N-acetyl-β-D-glucosaminyl 基团，然后去除保护基团而合成的。化合物 6a、6b 和 7a 经 N-乙酰基-β-D-葡糖胺苷酶水解后，在弱酸性速率测定条件（pH 值为 5.0）下，产物在长吸收波长（500、465 和 485 纳米）处显示出较高的吸光度。6a 和 7a 的 Km 值分别为 0.56 和 0.86 mM。在这些化合物中，7a 被认为是最有可能用于 N-乙酰基-β-D-葡糖胺酶速率测定的发色底物，因为它在酶水解作用下会产生从无色到橙色的明显颜色变化（λmax 280→485 nm），并且具有超过 30 mM 的较高水溶性。
• Enzymatic characterization of O-GlcNAcase isoforms using a fluorogenic GlcNAc substrate
  作者：Eun Ju Kim、Dae Ook Kang、Dona C. Love、John A. Hanover

  DOI：10.1016/j.carres.2006.03.004

  日期：2006.6

  A highly sensitive fluorogenic hexosaminidase substrate, fluorescein di(N-acetyl-beta-D-glucosaminide) (FDGlcNAc) was prepared essentially as described previously [Chem. Pharm. Bull. 1993, 41, 314] with some modifications. The fluorescent analog is a substrate for a number of hexosaminidases but here we have focused on the cytoplasmic O-GlcNAcase isoforms. Kinetic analysis using purified O-GlcNAcase and its splice variant (v-O-GlcNAcase) expressed in Escherichia coli suggests that FDGlcNAc is a much more efficient substrate (K-m = 84.9 mu M) than the conventional substrate, para-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside (pNP-beta-GlcNAc, K-m = 1.1 nM) and a previously developed fluorogenic substrate, 4-methylumbelliferyl 2-acetamido-2-deoxy-beta -D-glucopyranoside [MUGlcNAc, K-m = 0.43 mM; J. Biol. Chem. 2005, 280, 25313] for O-GlcNAcase. The variant O-GlcNAcase. a protein lacking the C-terminal third of the full-length O-GlcNAcase, exhibited a K-m of 2.1 mM with respect to FDGlcNAc. This shorter isoforni was not previously thought to exhibit O-GlcNAcase activity based on in vitro Studies with pNP-beta-GlcNAc. However, both O-GlcNAcase isoforms reduced O-GlcNAc protein levels extracted from HeLa and HT-29 cells in vitro. indicating that the splice variant is a bona fide O-GlcNAcase. Fluorescein di-N-acetyl-beta-D-galactosaminide (FDGalNAc) is not cleaved by these enzymes, consistent with previous findings that the O-GlcNAcase has substrate specificity toward O-GlcNAc but not O-GalNAc. The enzymatic activity of the shorter isoforni of O-GlcNAcase was first detected by using highly sensitive fluorogenic FDGlcNAc substrate. The finding that O-GlcNAcase exists as two distinct isoforms has a number of important implications for the role of O-GlcNAcase in hexosamine signaling. Published by Elsevier Ltd.