phytanyl-pyrophosphoryl-α-N,N′-diacetylchitobioside | 135137-89-2

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 异戊烯醇脂质
• 英文名称: phytanyl-pyrophosphoryl-α-N,N′-diacetylchitobioside
• 英文别名: PPGn2；phytanyl-pyrophosphate-GlcNAc2；[(2R,3R,4R,5S,6R)-3-acetamido-5-[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4-hydroxy-6-(hydroxymethyl)oxan-2-yl] [hydroxy(3,7,11,15-tetramethylhexadecoxy)phosphoryl] hydrogen phosphate
• CAS: 135137-89-2
• 化学式: C₃₆H₇₀N₂O₁₇P₂
• 分子量: 864.903
• InChiKey: IHJCWJQQTAUSHV-JYQKXATQSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  2.6
• 重原子数：
  57
• 可旋转键数：
  26
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.94
• 拓扑面积：
  289
• 氢给体数：
  9
• 氢受体数：
  17

反应信息

• 作为反应物：
  描述：

  phytanyl-pyrophosphoryl-α-N,N′-diacetylchitobioside 在 盐酸 、 乙二胺四乙酸 、 dithiothreitol 、 Helix pomatia β-mannosidase 、 recombinant His6-tagged S. cerevisiae Alg1 Δtransmembrane-spanning domain 、 magnesium chloride 、 NP-40 作用下， 以 甲醇 为溶剂， 反应 17.5h， 生成 N,N'-Diacetylchitobiose
  参考文献：
  名称：

  Quantitative study of yeast Alg1 beta-1, 4 mannosyltransferase activity, a key enzyme involved in protein N-glycosylation

  摘要：

  Background: Asparagine (N)-linked glycosylation begins with a stepwise synthesis of the dolichol-linked oligosaccharide (DLO) precursor, Glc3Man9GlcNAc2-PP-Dol, which is catalyzed by a series of endoplasmic reticulum membrane-associated glycosyltransferases. Yeast ALG1 (asparagine-linked glycosylation 1) encodes a beta-1, 4 mannosyltransferase that adds the first mannose onto GlcNAc2-PP-Dol to produce a core trisaccharide Man1GlcNAc2-PP-Dol.ALG1 is essential for yeast viability, and in humans mutations in the ALG1 cause congenital disorders of glycosylation known as ALG1-CDG. Alg1 is difficult to purify because of its low expression level and as a consequence, has not been well studied biochemically. Here we report a new method to purify recombinant Alg1 in high yield, and a mass spectral approach for accurately measuring its beta-1, 4 mannosyltransferase activity.Methods: N-terminally truncated yeast His-tagged Alg1 protein was expressed in Escherichia coli and purified by HisTrap HP affinity chromatography. In combination with LC-MS technology, we established a novel assay to accurately measure Alg1 enzyme activity. In this assay, a chemically synthesized dolichol-linked oligosaccharide analogue, phytanyl-pyrophosphoryl-alpha-N,N'-diacetylchitobioside (PPGn2), was used as the acceptor for the beta-1, 4 mannosyl transfer reaction.Results: Using purified Alg1, its biochemical characteristics were investigated, including the apparent K-m and V-max values for acceptor, optimal conditions of activity, and the specificity of its nucleotide sugar donor. Furthermore, the effect of ALG1-CDG mutations on enzyme activity was also measured.General significance: This work provides an efficient method for production of Alg1 and a new MS-based quantitative assay of its activity. (C) 2016 Elsevier B.V. All rights reserved.

  DOI：

  10.1016/j.bbagen.2016.09.023
• 作为产物：
  描述：

  2-acetamido-4-O-(2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl)-3,6-di-O-acetyl-2-deoxy-α-D-glucopyranosyl dibenzyl phosphate 在 palladium on activated charcoal 吡啶 、 三正丁胺 、 氢气 、 sodium methylate 、 silver(I) acetate 作用下， 以 甲醇 、 二氯甲烷 为溶剂， 反应 4.0h， 生成 phytanyl-pyrophosphoryl-α-N,N′-diacetylchitobioside
  参考文献：
  名称：

  Flitsch, Sabine L.; Pinches, Heather L.; Taylor, James P., Journal of the Chemical Society. Perkin transactions I, 1992, # 16, p. 2087 - 2094

  摘要：

  DOI：

文献信息

• Chemo-enzymatic synthesis of the ALG1-CDG biomarker and evaluation of its immunogenicity
  作者：Ji-Xiang Jia、Ndayambaje Yvan Kalisa、Tian-Tian Lu、Zhifang Zhou、Xiao-Dong Gao、Ning Wang

  DOI：10.1016/j.bmcl.2020.127614

  日期：2020.12

  Neu5Ac-α2, 6-Gal-β1, 4-GlcNAc-β1, 4-GlcNAc, was recently reported as the biomarker of ALG1-CDG, the disease caused by ALG1 deficiency. To develop a novel diagnostic method for ALG1-CDG, chemo-enzymatic synthesis of the tetrasaccharide biomarker linked to phytanyl phosphate and the biomarker’s immune stimulation were investigated in this study. The immunization study using liposomes bearing phytanyl-linked

  先天性糖基化疾病（CDG）是由与糖类生物合成途径有关的基因缺陷引起的增长中的疾病。最近报道了一种四糖，即Neu5Ac-α2、6-Gal-β1、4-GlcNAc-β1、4-GlcNAc，作为ALG1-CDG的生物标志物，该疾病是由ALG1缺乏引起的。为了开发一种新的ALG1-CDG诊断方法，本研究研究了与植烷磷酸酯连接的四糖生物标记物的化学酶法合成和生物标记物的免疫刺激作用。使用带有植烷酰基连接的四糖的脂质体进行的免疫研究表明，它们刺激了中等的免疫反应。诱导的抗体显示出对ALG1-CDG生物标志物的强结合特异性，表明其在医学应用中的潜力。
• Efficient Enzymatic Synthesis of the Core Trisaccharide ofN-Glycans with a Recombinantβ-Mannosyltransferase
  作者：Gregory M. Watt、Leigh Revers、Matthew C. Webberley、Iain B. H. Wilson、Sabine L. Flitsch

  DOI：10.1002/anie.199723541

  日期：1997.11.14
• Flitsch, Sabine L.; Pinches, Heather L.; Taylor, James P., Journal of the Chemical Society. Perkin transactions I, 1992, # 16, p. 2087 - 2094
  作者：Flitsch, Sabine L.、Pinches, Heather L.、Taylor, James P.、Turner, Nicholas J.

  DOI：——

  日期：——
• Quantitative study of yeast Alg1 beta-1, 4 mannosyltransferase activity, a key enzyme involved in protein N-glycosylation
  作者：Sheng-Tao Li、Ning Wang、Sha Xu、Jian Yin、Hideki Nakanishi、Neta Dean、Xiao-Dong Gao

  DOI：10.1016/j.bbagen.2016.09.023

  日期：2017.1

  Background: Asparagine (N)-linked glycosylation begins with a stepwise synthesis of the dolichol-linked oligosaccharide (DLO) precursor, Glc3Man9GlcNAc2-PP-Dol, which is catalyzed by a series of endoplasmic reticulum membrane-associated glycosyltransferases. Yeast ALG1 (asparagine-linked glycosylation 1) encodes a beta-1, 4 mannosyltransferase that adds the first mannose onto GlcNAc2-PP-Dol to produce a core trisaccharide Man1GlcNAc2-PP-Dol.ALG1 is essential for yeast viability, and in humans mutations in the ALG1 cause congenital disorders of glycosylation known as ALG1-CDG. Alg1 is difficult to purify because of its low expression level and as a consequence, has not been well studied biochemically. Here we report a new method to purify recombinant Alg1 in high yield, and a mass spectral approach for accurately measuring its beta-1, 4 mannosyltransferase activity.Methods: N-terminally truncated yeast His-tagged Alg1 protein was expressed in Escherichia coli and purified by HisTrap HP affinity chromatography. In combination with LC-MS technology, we established a novel assay to accurately measure Alg1 enzyme activity. In this assay, a chemically synthesized dolichol-linked oligosaccharide analogue, phytanyl-pyrophosphoryl-alpha-N,N'-diacetylchitobioside (PPGn2), was used as the acceptor for the beta-1, 4 mannosyl transfer reaction.Results: Using purified Alg1, its biochemical characteristics were investigated, including the apparent K-m and V-max values for acceptor, optimal conditions of activity, and the specificity of its nucleotide sugar donor. Furthermore, the effect of ALG1-CDG mutations on enzyme activity was also measured.General significance: This work provides an efficient method for production of Alg1 and a new MS-based quantitative assay of its activity. (C) 2016 Elsevier B.V. All rights reserved.