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Glycoproteomic analysis reveals the effects of bisecting GlcNAc in intrahepatic cholangiocarcinoma. 糖蛋白组学分析揭示了分割GlcNAc在肝内胆管癌中的作用。
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-12-01 DOI: 10.1007/s10719-022-10085-5
Wei Dan, Cheng Li, Jun Li, Pengfei Li, Miaomiao Xin, Zexuan Chen, Liuyi Dang, Zihao Yu, Jing Li, Jiechen Shen, Liangshuo Hu, Shisheng Sun

Intrahepatic cholangiocarcinoma (ICC) is the second major subtype of primary liver cancer and has caused more and more attention with increasing incidence and mortality worldwide. Our previous study found that bisecting N-glycans are commonly increased in ICC, while the effects and potential functions of bisecting GlcNAc in ICC are still largely unclear. In this study, we further confirmed that the structures of bisecting GlcNAc were significantly up-regulated in ICC compared with paracancer tissues by glycoproteomic data and lectin histochemistry. The expression of its glycosyltransferase MGAT3 was also up-regulated in ICC tissues at both mRNA and protein levels, and expression of MGAT3 is negatively correlated with overall survival explored by bioinformatic analyses and published datasets from 255 patients. Next, the silencing of MGAT3 could inhibit the growth and invasion of ICC cells, and overexpressing of MGAT3 only promoted ICC cell invasion. Further glycoproteomic analysis showed that the commonly glycoproteins modified by bisecting GlcNAc after MGAT3-overexpression in two ICC cell lines were mainly involved in cell movement-related biological processes, such as cell adhesion, integrin-related and ECM-receptor interaction. This study sheds light on the potential effects of bisecting GlcNAc in ICC cells and suggests that MGAT3 might be used as a potential target in the therapy of ICC.

肝内胆管癌(ICC)是原发性肝癌的第二大亚型,随着发病率和死亡率的上升,在世界范围内引起了越来越多的关注。我们之前的研究发现,在ICC中,等分n -聚糖普遍增加,而等分GlcNAc在ICC中的作用和潜在功能仍不清楚。在本研究中,我们通过糖蛋白组学数据和凝集素组织化学进一步证实,与癌旁组织相比,ICC中分切GlcNAc的结构显著上调。其糖基转移酶MGAT3的表达在ICC组织中的mRNA和蛋白水平也上调,通过生物信息学分析和来自255例患者的已发表数据集,MGAT3的表达与总生存率呈负相关。其次,沉默MGAT3可以抑制ICC细胞的生长和侵袭,过表达MGAT3只促进ICC细胞的侵袭。进一步的糖蛋白组学分析表明,在两种ICC细胞系中,mgat3过表达后,通过分割GlcNAc修饰的常见糖蛋白主要参与细胞运动相关的生物学过程,如细胞粘附、整合素相关和ecm受体相互作用。本研究揭示了分割GlcNAc在ICC细胞中的潜在作用,并提示MGAT3可能作为ICC治疗的潜在靶点。
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引用次数: 0
Effect of fermentation pH on the structure, rheological properties, and antioxidant activities of exopolysaccharides produced by Alteromonas australica QD. 发酵pH对澳大利亚异单胞菌胞外多糖结构、流变学特性及抗氧化活性的影响
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-12-01 DOI: 10.1007/s10719-022-10087-3
Fengshu Li, Xin Hu, Xiaojun Sun, Haoshuai Li, Jinren Lu, Yiming Li, Mutai Bao

The pH value was essential for the growth and metabolism of microorganisms. Acidic pH exopolysaccharide (AC-EPS) and alkaline pH exopolysaccharide (AL-EPS) secreted by A. australica QD mediated by pH were studied in this paper. The total carbohydrate content and molecular weight of AC-EPS (79.59% ± 2.24% (w/w), 8.374 × 105 Da) and AL-EPS (82.48% ± 1.46% (w/w), 6.182 × 105 Da) were estimated and compared. In AC-EPS, mannose (3.78%) and galactose (3.24%) content was more, while the proportion of glucuronic acid was less in comparison to AL-EPS. The scanning electron microscopy revealed the structural differences among the AC-EPS and AL-EPS. Thermogravimetric analysis showed degradation temperatures of 272.8 °C and 244.9 °C for AC-EPS and AL-EPS, respectively. AC-EPS was found to exhibit better rheological properties and emulsifying capabilities, while AL-EPS had superior antioxidant activities. Overall, both AC-EPS and AL-EPS have the potential to be used as emulsifiers and biological antioxidants.

pH值是微生物生长和代谢所必需的。研究了酸碱度介导的澳洲木质素QD分泌的酸性pH外多糖(AC-EPS)和碱性pH外多糖(AL-EPS)。比较了AC-EPS的总碳水化合物含量(79.59%±2.24% (w/w), 8.374 × 105 Da)和AL-EPS的总碳水化合物含量(82.48%±1.46% (w/w), 6.182 × 105 Da)。AC-EPS中甘露糖(3.78%)和半乳糖(3.24%)的含量高于AL-EPS,葡萄糖醛酸的含量低于AL-EPS。扫描电镜显示了AC-EPS和AL-EPS的结构差异。热重分析表明,AC-EPS和AL-EPS的降解温度分别为272.8℃和244.9℃。AC-EPS具有较好的流变性能和乳化性能,而AL-EPS具有较好的抗氧化活性。综上所述,AC-EPS和AL-EPS都具有作为乳化剂和生物抗氧化剂的潜力。
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引用次数: 2
A water-soluble polysaccharide from Eucommia folium: the structural characterization and anti-tumor activity in vivo. 杜仲叶水溶性多糖的结构表征及体内抗肿瘤活性。
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-12-01 DOI: 10.1007/s10719-022-10086-4
Zhi-Qian Yan, Su-Yun Ding, Pei Chen, Hui-Ping Liu, Meng-Li Chang, Shu-Yuan Shi

In this study, a water-soluble polysaccharide from Eucommia folium was extracted by hot water and purified using Sephadex G-200 gel columns. The results showed that the purified fraction (EFP) has a molecular weight of 9.98 × 105 Da and consisted of rhamnose, arabinose, galactose, glucose, mannose, xylose, galacturonic acid, and glucuronic acid (molar ratio: 0.226: 1.739: 2.183: 1: 0.155: 0.321: 0.358: 0.047). The combination of infrared spectroscopy and NMR analysis proved that EFP is an acidic polysaccharide whose main chain consists of α-L-Araf-(1 → , → 3,5)-α-Araf-(1 → , → 3)-β-Galp-(1 → , → 3,6)-β-Glcp-(1 → , → 2)-α-D-Manp-(1 → , → 4)-α-GalpA-(1 → , → 2,4)-α-Rhap-(1 → . In addition, the in vivo antitumoral activity of EFP was studied using a H22 tumor-bearing mice model. EFP effectively inhibited tumor growth in mice following intragastric administration. By Combining with the results of the apoptosis assay and JC-1 staining analysis, we confirmed that EFP induces apoptosis through the mitochondrial pathway. Furthermore, cell cycle analysis demonstrated that EFP blocks the cell cycle at S phase.

本研究采用热水提取杜仲叶水溶性多糖,并用Sephadex G-200凝胶柱进行纯化。结果表明,纯化后的EFP分子量为9.98 × 105 Da,由鼠李糖、阿拉伯糖、半乳糖、葡萄糖、甘露糖、木糖、半乳糖醛酸和葡萄糖醛酸组成,摩尔比为0.226:1.739:2.183:1:0.155:0.321:0.358:0.047。相结合的红外光谱和核磁共振分析证明了EFP是一种酸性多糖的主链由α-L-Araf -(1→3→5)-α-Araf -(1→→3)βgalp -(1→3→6)-β-Glcp -(1→→2)α-D-Manp -(1→→4)α-GalpA -(1→→2、4)-α世行-(1→。此外,采用H22荷瘤小鼠模型研究了EFP的体内抗肿瘤活性。EFP可有效抑制小鼠灌胃后的肿瘤生长。结合凋亡实验和JC-1染色分析结果,我们证实EFP通过线粒体途径诱导细胞凋亡。此外,细胞周期分析表明EFP在S期阻断细胞周期。
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引用次数: 3
Simple separation of glycosphingolipids in the lower phase of a Folch's partition from crude lipid fractions using zirconium dioxide. 使用二氧化锆从粗脂馏分中简单分离Folch分割下部相的鞘糖脂。
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-12-01 DOI: 10.1007/s10719-022-10080-w
Hideharu Nagasawa, Shota Miyazaki, Mamoru Kyogashima

A simple method was developed for the separation of glycosphingolipids (GSLs) from lipid mixtures, including phospholipids and cholesterol, using zirconium dioxide (zirconia, ZrO2). Although this procedure does not incorporate a mild alkali treatment, which is commonly used for eliminating glycerophospholipids, it can be used to remove both alkali-resistant sphingomyelin and glycerophospholipids possessing ether bonds. Importantly, when GSLs were dissolved in organic solvent together with cholesterol (Chol) and phospholipids, and loaded onto ZrO2, Chol did not bind to the ZrO2 but both the GSLs and phospholipids did. When eluted with 5 mg/mL of 2,5-dihydroxybenzoic acid in methanol, GSLs but not phospholipids were recovered, leaving the phospholipids bound to the ZrO2 particles. This method is particularly applicable for GSLs such as triglycosylceramides, tetraglycosylceramides and some pentaglycosylceramides, sulfatide and GM3 located in the lower phase of a Folch's partition, where significant amounts of phospholipids, Chol and neutral lipids reside along with GSLs. This method was successfully used to easily isolate GSLs from biological materials for their subsequent analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry with high resolution.

建立了一种用二氧化锆(ZrO2)从磷脂和胆固醇等脂质混合物中分离鞘糖脂(GSLs)的简便方法。虽然该方法不包括通常用于去除甘油磷脂的温和碱处理,但它可用于去除耐碱鞘磷脂和具有醚键的甘油磷脂。重要的是,当GSLs与胆固醇(Chol)和磷脂一起溶解在有机溶剂中,并装载到ZrO2上时,Chol不与ZrO2结合,但GSLs和磷脂都与ZrO2结合。当用5mg /mL 2,5-二羟基苯甲酸在甲醇中洗脱时,回收GSLs但不回收磷脂,使磷脂与ZrO2颗粒结合。该方法特别适用于位于Folch’s隔板下部相的gsl,如甘油三酯神经酰胺、四甘油三酯神经酰胺和一些五甘油三酯神经酰胺、硫脂和GM3,其中大量的磷脂、Chol和中性脂与gsl一起存在。该方法可方便地从生物材料中分离出GSLs,用于基质辅助激光解吸电离飞行时间质谱分析。
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引用次数: 1
A homogalacturonan from Lonicera japonica Thunb. disrupts angiogenesis via epidermal growth factor receptor and Delta-like 4 associated signaling. 取自金银花(Lonicera japonica Thunb)的一种同型半乳糖酸。通过表皮生长因子受体和δ样4相关信号干扰血管生成。
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-12-01 DOI: 10.1007/s10719-022-10088-2
Wenfeng Liao, Xiaodong Hu, Zhenyun Du, Peipei Wang, Kan Ding

A homogeneous polysaccharide named as LJW2F2 was extracted and purified from the flowers of Lonicera japonica Thunb. Structural characteristic indicated that LJW2F2 was a homogalacturonan composed of α-1,4-D-galacturonic acid with a molecular weight of 7.2 kDa. Previous investigation suggested that homogalacturonan might impede angiogenesis, however the mechanism is still vague. Here we reported that LJW2F2 significantly disrupted capillary-like tube formation of human microvascular endothelia cells (HMEC-1) on matrigel as well as the cells migration. Mechanism study revealed that LJW2F2 might inactivate phosphorylation of epidermal growth factor receptor (EGFR), subsequently suppress Raf, mitogen-activated protein kinase (MEK) and extracellular-related kinase (ERK) phosphorylation. Moreover, LJW2F2 markedly decreased the expression of Notch1 and Delta-like ligand 4 (Dll4). Therefore, our results suggested that LJW2F2 might be a potential angiogenesis inhibitor via disturbing multiple signaling pathways.

从金银花中提取纯化了一种均相多糖LJW2F2。结构特征表明LJW2F2为α-1,4- d -半乳糖醛酸组成的均半乳糖醛酸,分子量为7.2 kDa。以往的研究表明,均半乳糖酸可能阻碍血管生成,但其机制尚不清楚。本研究报道LJW2F2显著破坏基质上人微血管内皮细胞(HMEC-1)的毛细血管样管形成和细胞迁移。机制研究表明,LJW2F2可能使表皮生长因子受体(EGFR)磷酸化失活,进而抑制Raf、丝裂原活化蛋白激酶(MEK)和细胞外相关激酶(ERK)磷酸化。LJW2F2显著降低Notch1和δ样配体4 (Dll4)的表达。因此,我们的研究结果表明LJW2F2可能是一种潜在的血管生成抑制剂,通过干扰多种信号通路。
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引用次数: 2
Thrombospondin type 1 repeat-derived C-mannosylated peptide attenuates synaptogenesis of cortical neurons induced by primary astrocytes via TGF-β. 血小板反应蛋白1型重复源性c -甘露糖基化肽通过TGF-β减弱原代星形胶质细胞诱导的皮质神经元突触发生。
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-10-01 Epub Date: 2021-11-18 DOI: 10.1007/s10719-021-10030-y
Kazuchika Nishitsuji, Midori Ikezaki, Shino Manabe, Kenji Uchimura, Yukishige Ito, Yoshito Ihara

C-Mannosylation is a rare type of protein glycosylation and is reportedly critical for the proper folding and secretion of parental proteins. Still, the effects of C-mannosylation on the biological functions of these modified proteins remain to be elucidated. The Trp-x-x-Trp (WxxW) sequences, whose first tryptophan (Trp) can be C-mannosylated, constitute the consensus motifs for this glycosylation modification and are commonly found in thrombospondin type 1 repeats that regulate molecular functions of thrombospondin 1 in binding and activation of transforming growth factor β (TGF-β). TGF-β plays critical roles in the control of the central nervous system including synaptogenesis. Here, we investigated whether C-mannosylation of the synthetic Trp-Ser-Pro-Trp (WSPW) peptide may confer certain functions to this peptide in TGF-β-mediated synaptogenesis. By using primary cultured rat astrocytes and cortical neurons, we found that the C-mannosylated WSPW (C-Man-WSPW) peptide, but not non-mannosylated WSPW peptide, suppressed astrocyte-conditioned medium (ACM)-stimulated synaptogenesis. C-Man-WSPW peptide inhibited both ACM- and recombinant mature TGF-β1-induced activations of Smad 2, an important mediator in TGF-β signaling. Interactions of recombinant mature TGF-β with the C-Man-WSPW peptide were similar to those with non-C-mannosylated WSPW peptide. Taken together, our results reveal a novel function of C-mannosylation of the WxxW motif in signaling and synaptogenesis mediated by TGF-β. Molecular details of how C-mannosylation affects the biological functions of WxxW motifs deserve future study for clarification.

c -甘露糖基化是一种罕见的蛋白质糖基化,据报道对亲本蛋白的正常折叠和分泌至关重要。然而,c -甘露糖基化对这些修饰蛋白的生物学功能的影响仍有待阐明。Trp-x-x-Trp (WxxW)序列,其第一个色氨酸(Trp)可以被c -甘露糖基化,构成了这种糖基化修饰的共识基序,并且通常存在于调节血小板反应蛋白1结合和激活转化生长因子β (TGF-β)的分子功能的血小板反应蛋白1重复序列中。TGF-β在中枢神经系统包括突触发生的控制中起关键作用。在这里,我们研究了c -甘露糖基化合成的Trp-Ser-Pro-Trp (WSPW)肽是否在TGF-β介导的突触发生中赋予该肽某些功能。通过原代培养的大鼠星形胶质细胞和皮质神经元,我们发现c -甘露糖化的WSPW (C-Man-WSPW)肽,而非甘露糖化的WSPW肽,抑制星形胶质细胞条件培养基(ACM)刺激的突触发生。C-Man-WSPW肽抑制ACM-和重组成熟TGF-β1诱导的Smad 2的激活,Smad 2是TGF-β信号传导的重要介质。重组成熟TGF-β与C-Man-WSPW肽的相互作用类似于与非c -甘露糖基化WSPW肽的相互作用。综上所述,我们的研究结果揭示了WxxW基序c -甘露糖基化在TGF-β介导的信号传导和突触发生中的新功能。c -甘露糖基化如何影响WxxW基序的生物学功能的分子细节值得进一步研究以澄清。
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引用次数: 1
Roland Schauer – Obituary Roland Schauer–讣告
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-08-03 DOI: 10.1007/s10719-022-10070-y
Anthony Corfield, J. Kamerling
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引用次数: 0
Correction to: Role of sialylated glycans on bovine lactoferrin against influenza virus 更正:唾液酸化聚糖对牛乳铁蛋白对抗流感病毒的作用
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-06-10 DOI: 10.1007/s10719-022-10071-x
Xilong Wang, Lixin Yue, Liuyi Dang, Jiajun Yang, Zhuo Chen, Xiurong Wang, Jian Shu, Zheng Li
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引用次数: 0
Sialidase NEU3 and its pathological significance 唾液酸酯酶NEU3及其病理意义
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-06-08 DOI: 10.1007/s10719-022-10067-7
T. Miyagi, Koji Yamamoto
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引用次数: 8
Structural analysis and in vitro antitumor effect of polysaccharides from Pholiota adiposa 脂肪木多糖的结构分析及体外抗肿瘤作用
IF 3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2022-06-08 DOI: 10.1007/s10719-022-10065-9
Jiao Zhou, Jinhua Gong, Yangyang Chai, D. Li, Cong Zhou, Chuansheng Sun, J. Regenstein
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引用次数: 2
期刊
Glycoconjugate Journal
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