ENZYME AND MICROBIAL TECHNOLOGY
·
2025
·
IF 3.7
Chen-Xin Guo,Jiyan Li,Yu Wang,Tang Li,Heng Yin
Stevia rebaudiana , a perennial herb, is recognized not only for its sweet steviol glycosides but also for its rich flavonoid content, which confer pharmacological properties including anti-inflammatory, antimicrobial, and anticancer activities. However, the enzymatic basis underlying flavonoid modification in S. rebaudiana remains poorly understood. In this study, we identified, cloned, and heterologously expressed a novel flavonoid glycosyltransferase gene, SrUGT72B1 in E. coli . The recombinant SrUGT72B1 catalyzed the glycosylation of multiple flavonoids using UDP-glucose as the primary sugar donor, and exhibited broad substrate promiscuity toward apigenin, luteolin, phloretin and kaempferol. In addition to UDP-glucose, SrUGT72B1 also accepted UDP-xylose and UDP-rhamnose, with UDP-glucose exhibiting the highest catalytic efficiency. Biochemical characterization revealed that the enzyme functions optimally at pH 9.0 and 50 °C. Notably, SrUGT72B1 demonstrates regioselective 5-O-glycosylation toward apigenin, a rare activity among plant glycosyltransferases. Molecular docking and molecular dynamics simulations provided structural insights into this unique regioselectivity and substrate recognition. Together, these findings establish SrUGT72B1 as a previously uncharacterized flavonoid 5-O-glycosyltransferase, expanding the functional landscape of plant UGTs and offering potential applications in the biosynthesis of value-added flavonoid glycosides.
研究领域:
植物学
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生物化学
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分子生物学
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药理学
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PubMed
高影响因子
Nature Communications
·
2025
·Vol. 16
·期 1
·1-17
·
IF 15.7
Liu Qiannan,Lu Xiaoyan,Deng Yao,Zhang Han,Wei Rumeng,Li Hongrui,Feng Ying,Wei Juan,Ma Fang,Zhang Yan,Zou Xia
Protein O-glycosylation plays critical roles in sperm formation and maturation. However, detailed knowledge on the mechanisms involved is limited due to lacking characterization of O-glycoproteome of testicular germ cells. Here, we performed a systematic analysis of site-specific O-glycosylation in mouse testis, and established an O-glycoproteome map with 349 O-glycoproteins and 799 unambiguous O-glycosites. Moreover, we comprehensively investigated the distribution properties of O-glycosylation in testis and identified a region near the N-terminal of peptidase S1 domain that is susceptible to O-glycosylation. Interestingly, we found dynamic changes with an increase Tn and a decrease T structure from early to mature developmental stages. Notably, the importance of O-glycosylation was supported by its effects on the stability, cleavage, and interaction of acrosomal proteins. Collectively, these data illustrate the global properties of O-glycosylation in testis, providing insights and resources for future functional studies targeting O-glycosylation dysregulation in male infertility.
研究领域:
生殖生物学
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蛋白质组学
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分子生物学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2025
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IF 6.7
Zhongyu Wang,Yan Wang,Taiheng Ma,He Zhu,Deguang Sun,Zhimou Guo,Xinmiao Liang,Mingliang Ye,Mingming Dong
Tn (GalNAcα1-O-S/T) and sTn (Neu5Acα2-6GalNAcα1-O-S/T) antigens, the immature O-glycans, are strongly correlated with cancer aggressiveness. Tn and sTn antigens exhibit significant differences between patients and healthy people, and they have emerged as potential biomarkers for cancer diagnosis and prognosis. Even though several chemical or chemoenzymatic labeling methods have been developed, the simultaneous analysis of Tn and sTn glycosites at the proteome level still faces great challenges. Here, we propose a label-free method for the sensitive identification of Tn/sTn glycosites from complex proteome samples. Because Tn/sTn antigens often densely occur on protein regions rich in Ser/Thr residues but lacking Lys/Arg residues, the trypsin digestion of these O-glycoproteins will generate relatively large peptides carrying multiple O-glycans. In this study, large tryptic O-glycopeptides were specifically enriched using hydrophilic interaction liquid chromatography. Subsequently, glycopeptides containing a single Tn/sTn antigen were generated by N-terminal-specific protease digestion, enabling the identification of Tn/sTn glycosites at the peptide N-termini. We demonstrated the general applicability of this method to analyze challenging samples, including HeLa cells and pancreatic ductal adenocarcinoma serum, where Tn/sTn are low-abundance O-glycans. Furthermore, a total of 191 Tn/sTn glycosites were identified on 200 glycopeptides from 136 proteins across two pancreatic cancer cell lines and serum. In summary, the method is broadly applicable, sensitive and well fitted to analyze a large cohort of clinical samples.
研究领域:
生物化学
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分子生物学
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肿瘤学
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蛋白质组学
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DOI
PubMed
高影响因子
Foods
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2025
·Vol. 14
·期 15
·2708
·
IF 5.1
Guanghui Li,Kit-Leong Cheong,Yunhua He,Ahluk Liew,Jiaxuan Huang,Chen Huang,Saiyi Zhong,Malairaj Sathuvan
AlthoughHylocereus polyrhizuspulp residues polysaccharides (HPPP) have shown potential in improving metabolic disorders and intestinal barrier function, the mechanism by which they exert their effects through regulating O-glycosylation modifications in the mucus layer remains unclear. Therefore, this study established a HFD-induced obese colitis mouse model (n= 5 per group) and combined nano-capillary liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) technology to quantitatively analyze the dynamic changes in O-glycosylation. Additionally, through quantitative O-glycosylation proteomics and whole-proteome analysis, we identified 155 specifically altered O-glycosylation sites in colon tissue, with the glycosylation modification level of the MUC2 core protein increased by approximately 2.1-fold. The results indicate that HPPP alleviates colonic mucosal damage by regulating interactions between mucus O-glycosylation. Overall, we demonstrated that HPPP increases HFD-induced O-glycosylation sites, improves intestinal mucosal structure in obese mice, and provides protective effects against obesity-induced intestinal mucosal damage.
研究领域:
营养科学
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生物化学
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分子生物学
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食品科学
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胃肠病学
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DOI
PubMed
高影响因子
Oxidative Medicine and Cellular Longevity
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2022
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IF 7.31
Chang-Yu Yan,Jie Sun,Gui-Yuan Yu,Jiang-Han-Zi Liu,Rong-Ping Huang,Shao-Cong Han,Qiong-Yi Zhang,Xiao-Min Li,Jian-Gang Yan,Hiroshi Kurihara,Wei-Xi Li,Yi-Fang Li,Rong-Rong He
Neural tube defect (NTD) is the most common and severe embryopathy causing embryonic malformation and even death associated with gestational diabetes mellitus (GDM). Leu-Pro-Phe (LPF) is an antioxidative tripeptide isolated from hydrolysates of corn protein. However, the biological activity of LPF in vivo and in vitro remains unclear. This study is aimed at investigating the protective effects of tripeptide LPF against NTD in the high glucose exposure condition and delineate the underlying biological mechanism. We found that LPF alleviated NTD in the high glucose-exposed chicken embryo model. In addition, DF-1 chicken embryo fibroblast was loaded with high glucose for induction of oxidative stress and abnormal O-GlcNAcylation in vitro. LPF significantly decreased accumulation of reactive oxygen species and content of malondialdehyde in DF-1 cells but increased the ratio of reduced glutathione and oxidized glutathione in chick embryo. Oxygen radical absorbance capacity results showed that LPF itself had good free radical scavenging capacity and could enhance antioxidant activity of the cell content. Mechanistic studies suggested that the resistance of LPF to oxidative damage may be related to promotion of NRF2 expression and nuclear translocation. LPF alleviated the overall O-GlcNAcylation level of cellular proteins under high glucose conditions and restored the level of Pax3 protein. Collectively, our findings indicate that LPF peptide could act as a nutritional supplement for the protection of development of embryonic neural tube affected by GDM.
研究领域:
营养学
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胚胎学
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分子生物学
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生物化学
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DOI
PubMed
高影响因子
JOURNAL OF NUTRITIONAL BIOCHEMISTRY
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2022
·
IF 6.117
Peiheng Han,Yujuan Yu,Li Zhang,Zheng Ruan
Citrus peel is rich in bioactive components, especially polyphenols, which are considered to have great potential in the prevention of intestinal diseases . The intestinal mucus barrier is the first defense against the invasion of foreign substances. In this study, we aimed to explore the possibility and mechanism of citrus peel in alleviating the mucus barrier damage in high-fat-diet (HFD) mice. We found that citrus peel powder (CPP) supplementation effectively reduced body weight, fat weight, intestinal permeability , hyperlipidemia , and systemic inflammation in HFD-fed mice. In particular, CPP increased the number of goblet cells , the protein expression of Mucin-2 (Muc2), and the thickness of the mucus layer, thereby strengthening the colonic mucus barrier function. Moreover, CPP supplementation also reduced the expression of endoplasmic reticulum stress (ERS) proteins (GRP78 and CHOP) and increased the expression of T-synthase ( O- glycosylation rate-limiting enzyme) and its chaperone protein (Cosmc) in the colon of HFD-fed mice, which suggested that CPP could improve the abnormal protein folding and O -glycosylation of Muc2 during processing and modification. In summary, our study indicates that CPP plays an effective role in relieving mucus barrier damage by improving the production and properties of Muc2, providing new perspectives on the development of CPP as a dietary supplement for strengthening the intestinal barrier.
研究领域:
营养学
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胃肠病学
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分子生物学
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药理学
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DOI
PubMed
高影响因子
CANCER SCIENCE
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2024
·
IF 4.5
Luhaoran Sun,Zeyu Li,Peng Shu,Min Lu
High expression of truncated O-glycans Tn antigen predicts adverse clinical outcome in patients with clear cell renal cell carcinoma (ccRCC). To understand the biosynthetic underpinnings of Tn antigen changes in ccRCC, we focused on N-acetylgalactosaminyltransferases (GALNTs, also known as GalNAcTs) known to be involved in Tn antigen synthesis. Data from GSE15641 profile and local cohort showed that GALNT6 was significantly upregulated in ccRCC tissues. The current study aimed to determine the role of GALNT6 in ccRCC, and whether GALNT6-mediated O-glycosylation aggravates malignant behaviors. Gain- and loss-of-function experiments showed that overexpression of GALNT6 accelerated ccRCC cell proliferation, migration, and invasion, as well as promoted ccRCC-derived xenograft tumor growth and lung metastasis. In line with this, silencing of GALNT6 yielded the opposite results. Mechanically, high expression of GALNT6 led to the accumulation of Tn antigen in ccRCC cells. By undertaking immunoprecipitation coupled with liquid chromatography/mass spectrometry, vicia villosa agglutinin blot, and site-directed mutagenesis assays, we found that O-glycosylation of prohibitin 2 (PHB2) at Ser161 was required for the GALNT6-induced ccRCC cell proliferation, migration, and invasion. Additionally, we identified lens epithelium-derived growth factor (LEDGF) as a key regulator of GALNT6 transcriptional induction in ccRCC growth and an upstream contributor to ccRCC aggressive behavior. Collectively, our findings indicate that GALNT6-mediated abnormal O-glycosylation promotes ccRCC progression, which provides a potential therapeutic target in ccRCC development.
研究领域:
癌症生物学
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分子生物学
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细胞生物学
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肿瘤学
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产品关联
DOI
PubMed
高影响因子
npj Biofilms and Microbiomes
·
2023
·Vol. 9
·期 1
·1-13
·
IF 9.2
Zhao Tong,Zhang Yue,Nan Linhua,Zhu Qing,Wang Shukai,Xie Yutao,Dong Xinling,Cao Cui,Lin Xiaoliang,Lu Yu,Liu Yuxia,Huang Linjuan,Gong Guiping,Wang Zhongfu
Understanding how dietary polysaccharides affect mucin O -glycosylation and gut microbiota could provide various nutrition-based treatments. Here, the O -glycan profile of the colonic mucosa and gut microbiome were investigated in C57BL/6J mice fed six structurally diverse dietary polysaccharides and a mixture of six fibers. Dietary polysaccharides increased total O -glycans, mainly by stimulating neutral glycans. Highly branched arabinogalactan promoted terminally fucosylated core 1 O -glycans; whereas linear polysaccharides, including pectin, konjac glucomannan, inulin, and the fiber mixture, favored terminally di-fucosylated O -glycans. The last three polysaccharides also lowered the level of sulfated O -glycans and sialylated mono-fucosylated O -glycans. Varied monosaccharide composition in mixed polysaccharides had a synergistic beneficial effect, boosting fucosylated neutral glycans, decreasing acidic glycans, and stimulating microbial richness and diversity. Dietary polysaccharides containing arabinose and sulfate groups enhanced the relative abundances of Akkermansia and Muribaculaceae , respectively. The present comparison reveals the relationship between dietary polysaccharide structure, mucin O -glycan composition, and intestinal microorganisms.
研究领域:
营养学
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胃肠病学
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微生物学
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生物化学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Shuang Yue,Xiaotong Wang,Lei Wang,Jiajia Li,Yufeng Zhou,Yan Chen,Zeyang Zhou,Xiaodong Yang,Xiaofeng Shi,Song Gao,Zhongmin Wen,Xiaojun Zhu,Yan Wang,Shuang Yang
The Tn antigen, an immature truncated O-glycosylation, is a promising biomarker for cancer detection and diagnosis. However, reliable methods for analyzing O-GalNAcylation and complex O-glycosylation are lacking. Here, we develop a novel method, MOTAI, for the sequential analysis of O-glycosylation using different O-glycoproteases. MOTAI conjugates glycopeptides on a solid support and releases different types of O-glycosylation through sequential enzymatic digestion by O-glycoproteases, including OpeRATOR and IMPa. Because OpeRATOR has less activity on O-GalNAcylation, MOTAI enriches O-GalNAcylation for subsequent analysis. We demonstrate the effectiveness of MOTAI by analyzing fetuin O-glycosylation and Jurkat cell lines. We then apply MOTAI to analyze colorectal cancer and benign colorectal polyps. We identify 32 Tn/sTn-glycoproteins and 43 T/sT-glycoproteins that are significantly increased in tumor tissues. Gene Ontology analysis reveals that most of these proteins are ECM proteins involved in the adhesion process of the intercellular matrix. Additionally, the protein disulfide isomerase CRELD2 has a significant difference in Tn expression, and the abnormally glycosylated T345 and S349 O-glycosylation sites in cancer group samples may promote the secretion of CRELD2 and ultimately tumorigenesis through ECM reshaping. In summary, MOTAI provides a powerful new tool for the in-depth analysis of O-GalNAcylation and complex O-glycosylation. It also reveals the upregulation of Tn/sTn-glycoproteins in colorectal cancer, which may provide new insights into cancer biology and biomarker discovery.
研究领域:
癌症生物学
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糖生物学
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生物化学
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分子生物学
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临床诊断
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Tong Zhao,Sining Liu,Yutong Shuai,Xinyi Zhang,Min Chen,Sijie Pei,Yuxi Duan,Shukai Wang,Yu Lu,Zhongfu Wang,Guiping Gong,Linjuan Huang
Pectin is a structurally complex heteropolysaccharide that affects intestinal microorganisms and mucin O -glycans. The present study employed an in vitro model to investigate dynamic changes in microbiota during pectin fermentation. Residual pectin fragments arising from its fermentation were applied to HT29-MTX cells to study the effect of pectin structure on mucin O -glycosylation. Prevotella , Bacteroides , and Parabacteroides were found to preferentially degrade galactose, arabinose, and on the rhamnogalacturonan RG-I side chain region and methyl esterification groups of pectin. Bifidobacterium , Enterococcus , Megamonas , and Dorea metabolized the galacturonic HG region on pectin to produce butyrate. All pectin fragments were found to up-regulate total O -glycans (1.55–2.73 fold) and neutral O -glycans (1.11–1.49 fold) on HT29-MTX mucins. The large HG fragment (81.04 kDa) increased significantly the amount of non-fucosylated glycans (by 2.46-fold); whereas the small HG fragment (16.02 kDa) promoted fucosylated (by 9.25 fold), and especially di-fucosylated O -glycans. Collectively, these results demonstrate that gut microorganisms degrade pectin fragments in the following order of utilization: RG-I, RG-II, and HG. The small fragment of HG improves the expression of fucosylated O -glycans in HT29-MTX cells, mainly owing to an increase in di-fucosylated O -glycans.
研究领域:
微生物学
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胃肠病学
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细胞生物学
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营养科学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）