TALANTA
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2020
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IF 5.339
Nan Yin,Ruihuan Zhao,Jie Zhang,Dingding Yang,Zhimin Guo,Rui Liu,Xin Yao
Abnormal O-linked N-acetylglucosamine (O-GlcNAc) concentrations have been associated with many diseases, but the lack of accurate detection method limited O-GlcNAc to be used as a biomarker in clinical diagnosis. Then O-GlcNAc transferase (OGT) has drawn researchers’ attention as it closed related to the level of O-GlcNAc and be considered to be a promising new target for diseases diagnosis. Nevertheless, the existing OGT detection methods are either need labeling or the sensitity can not meet the needs of clinic testing. Herein, a label-free and sensitive SPR biosensor was developed for accurate detection of OGT based on a multi-functional peptide. The designed peptide contains three recognition sites, one is the cleavage site of protease K, one is the O-GlcNAcylated site by OGT, and another is six histidine which be used as the signal report probe to recognize Ni 2+ . The immobilized peptide would be cleavaged by proteinase K, then the His-tag residue part will leave the surface of Au film, resulting less His-tag could bind to Ni 2+ and a small SPR signal would be record. If the peptide is O-GlcNAcylated by OGT, the cleaving reaction would be limited due to the adjacent site of O-GlcNAcylation. Then more His-tag can be left on the Au film and a bigger SPR signal could be record, this signal is associated with the concentration of OGT. Utilizing the change of the peptide configuration as a signal report probe for OGT detection not only avoids labeling of peptide, but also makes the method more sensitive. The determination linear range of OGT is from 2.00 × 10 −13 to 5.00 × 10 −8 M with a detection limit of 1.19 × 10 −13 M, and the separation of two enzyme reactions ensured the high selectivity of the method. Finally, the sensing system was successfully used for OGT detection in blood samples with satisfied recovery. In summary, the label-free SPR platform for accurate detection of OGT in real samples is helpful to promote OGT serve as a biomarker for early clinical diagnosis of O-GlcNAc related diseases.
研究领域:
生物化学
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分析化学
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生物传感器
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临床诊断
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PubMed
高影响因子
MOLECULES
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2018
·Vol. 23
·期 7
·1649
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IF 3.098
Qi Cheng,Yalu Cui,Nao Xiao,Jishun Lu,Chen-Jie Fang
TheO-linked β-N-acetylglucosamine (O-GlcNAc) modification is an abundant post-translational modification in eukaryotic cells, which plays a fundamental role in the activity of many cells and is associated with pathologies like type II diabetes, Alzheimer’s disease or some cancers. However, the precise connexion betweenO-GlcNAc-modified proteins and their function in cells is largely undefined for most cases. Confocal microscopy is a powerful and effective tool for in-cell elucidation of the function of biological molecules. Chemical labeling of non-ultraviolet or non-fluorescent carbohydrates with fluorescent tag is an essential step that makes intra-cellular microscopic inspection possible. Here we report a strategy based on the 1,3-dipolar cycloaddition, called click chemistry, between unnaturalN-acetylglucosamine (GlcNAc) analogues Ac4GlcNAc (substituted with an azido group) and the corresponding fluorescent tag Ru(bpy)2(Phen-alkyne)Cl2(4) to synthesize the fluorescent dye Ru(bpy)2(Phen-Ac4GlcNAc)Cl2(5) under mild and neutral reaction conditions. Moreover,5showed good stability, desirable fluorescence characteristics, and exhibited rather low levels of cytotoxicity against sensitive MCF-7 cells. Additionally, we have achieved successful fluorescent imaging of5transported in living MCF-7 cells. Cell images displayed that proteins are potentially labelled with5in the cytoplasm.Keywords:O-GlcNAc;click chemistry;cell imaging;fluorescent labeling;living cells
研究领域:
生物化学
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分子生物学
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细胞生物学
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化学
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高影响因子
Nature Communications
·
2024
·Vol. 15
·期 1
·1-16
·
IF 14.7
Cao Shiyang,Wang Tong,Ren Yifan,Wu Gengshan,Zhang Yuan,Tan Yafang,Zhou Yazhou,Chen Hongyan,Zhang Yu,Song Yajun,Yang Ruifu,Du Zongmin
Post-translational addition of O-linked N-acetylglucosamine (O-GlcNAc) to proteins is commonly associated with a variety of stress responses and cellular processes in eukaryotes, but its potential roles in bacteria are unclear. Here, we show that protein HmwC acts as an O-GlcNAc transferase (OGT) responsible for O-GlcNAcylation of multiple proteins in Yersinia pestis , a flea-borne pathogen responsible for plague. We identify 64 O-GlcNAcylated proteins (comprising 65 sites) with differential abundance under conditions mimicking the mammalian host (Mh) and flea vector (Fv) environments. Deletion of hmwC , encoding a putative OGT, structurally distinct from any existing member of the GT41 family, results in reduced O-GlcNAcylation, reduced growth, and alterations in virulence properties and survival under stress. Purified HmwC can modify target proteins in vitro using UDP-GlcNAc as sugar donor. One of the target proteins, OsdY, promotes Y. pestis survival under oxidative stress conditions. Thus, our results support that regulation of antioxidative responses through O-GlcNAcylation may be a conserved process shared by prokaryotes and eukaryotes.
研究领域:
微生物学
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病理学
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分子生物学
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传染病学
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高影响因子
MOLECULAR PHARMACEUTICS
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2023
·
IF 4.9
Bingyi Lin,Siyuan Chai,Qijun Zhang,Yuejie Lu,Jiahao Hu,Jie Zhang,Yong-Zhong Du,Liming Wu
O-linked-N-acetylglucosaminylation (O-GlcNAcylation) plays a key role in hepatocellular carcinoma (HCC) development, and the inhibition of O-GlcNAcylation has therapeutic potential. To decrease the systemic adverse events and increase targeting, we used sialic acid (SA)-decorated liposomes loaded with OSMI-1, an inhibitor of the O-GlcNAcylation, to further improve the anti-HCC effect. Fifty pairs of HCC tissue samples and the cancer genome atlas database were used to analyze the expression of O-GlcNAc transferase (OGT) and its effects on prognosis and immune cell infiltration. OSMI-1 cells were treated with SA and liposomes. Western blotting, immunofluorescence, cell proliferation assay, flow cytometry, enzyme-linked immunosorbent assay, immunohistochemistry, and tumorigenicity assays were used to investigate the antitumor effect of SA-modified OSMI-1 liposomes in vitro and in vivo. OGT was highly expressed in HCC tissues, negatively correlated with the degree of tumor infiltration of CD8+ and CD4+T cells and prognosis, and positively correlated with the degree of Treg cell infiltration. SA-modified OSMI-1 liposome (OSMI-1-SAL) was synthesized with stable hydrodynamic size distribution. Both in vitro and in vivo, OSMI-1-SAL exhibited satisfactory biosafety and rapid uptake by HCC cells. Compared to free OSMI-1, OSMI-1-SAL had a stronger capacity for suppressing the proliferation and promoting the apoptosis of HCC cells. Moreover, OSMI-1-SAL effectively inhibited tumor initiation and development in mice. OSMI-1-SAL also promoted the release of damage-associated molecular patterns, including anticalreticulin, high-mobility-group protein B1, and adenosine triphosphate, from HCC cells and further promoted the activation and proliferation of the CD8+ and CD4+T cells. In conclusion, the OSMI-1-SAL synthesized in this study can target HCC cells, inhibit tumor proliferation, induce tumor immunogenic cell death, enhance tumor immunogenicity, and promote antitumor immune responses, which has the potential for clinical application in the future.
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高影响因子
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
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2026
·
IF 2.7
Qiushi Chen,Wangzhirui Cheng,Can Li,Yi Man Eva Fung,Jiaqi Wang,Han Liu,Xuechen Li
Protein glycosylation plays essential roles in various biological processes, and thus determining the glycan structure present on the protein is essential to comprehensively understand these events. However, distinguishing saccharide stereoisomers is challenging, especially when their structures are very similar and their molecular weight and potential glycosylation sites are identical. One representative example is O-linked β-N-acetylglucosamine (O-GlcNAc) and O-linked α-N-acetylgalactosamine (Tn antigen). Traditional biochemistry approaches used in separating O-GlcNAc- and Tn antigen-modified peptides mainly include chemical derivatizations, lectins, and antibodies. However, subsequent mass spectrometry (MS) analysis is still required if one aims to determine the exact glycosylation site. Herein, a straightforward approach using the ratio of relative abundance (RA) of two fragment ions (RA126.055/RA138.055) in higher-energy collisional dissociation (HCD) MS without relying on the traditional biochemistry technique is reported to discriminate between O-GlcNAc and Tn antigen. This ratio was verified by synthetic glycopeptides and proteomic analysis in HeLa cells, where 10 proteins were found to be O-GlcNAcylation and 4 proteins were found to be Tn antigen-modified. Overall, this method can be extensively employed in liquid chromatography–mass spectrometry (LC-MS)-based proteomic studies and thus is of importance in biological and biomedical research.
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高影响因子
Advanced Science
·
2026
·
IF 14.1
Duoli Xie,Jie Huang,Chunhao Cao,Zhenchao Tang,Xingchen Li,Jianmin Guo,Xinxin Chen,Fang Wang,Dongyi He,Jie Li,Yu Du,Bing He,Jianhua Yao,Aiping Lu,Chao Liang
Rheumatoid arthritis (RA) progression is driven by the pathogenic transformation of stromal fibroblast‐like synoviocytes (FLSs). Using artificial intelligence‐guided analysis of a synovial single‐cell transcriptomic dataset, we identify O‐GlcNAc transferase (OGT) as a key regulator enriched in RA‐FLSs. Gain‐ and loss‐of‐function studies demonstrate that OGT is necessary to drive aggressive FLS phenotypes and exacerbate experimental arthritis. Mechanistically, OGT‐mediated O‐GlcNAcylation stabilizes SAP130, promoting histone deacetylation at the BTG2 promoter. This facilitates the deposition of repressive histone methylation marks, leading to BTG2 epigenetic silencing and ultimately fueling FLS aggression. To translate this mechanistic insight, we develop an FLS‐targeted proteolysis‐targeting chimera (PROTAC) by conjugating the OGT inhibitor OSMI‐1 to the AS1411 aptamer, which selectively binds surface nucleolin (NCL) overexpressed on pathogenic FLSs and, upon internalization, employs NCL as a molecular bridge to recruit the E3 ligase MDM2 for cell‐selective OGT degradation. This PROTAC suppresses FLS pathogenicity, attenuates arthritis in mice, and demonstrates additive therapeutic benefit when combined with the TNF‐α inhibitor etanercept. Collectively, our work establishes a translational paradigm that progresses from AI‐driven target discovery and mechanistic elucidation to the rational design of a cell‐type‐specific degradation therapy, offering a strategy to overcome the stromal‐driven therapeutic barrier in RA. AI‑guided single‑cell transcriptomics identifies OGT as a key regulator enriched in rheumatoid arthritis fibroblast‑like synoviocytes (RA‐FLSs). Inflammatory stimuli activate the OGT–SAP130 axis, which modulates the H3K27ac/H3K27me3 balance and epigenetically silences BTG2, promoting aggressive transformation of RA‑FLSs. A targeted PROTAC is developed to selectively degrade OGT in pathogenic FLSs, suppressing their aggressive phenotypes for RA therapy.
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高影响因子
CHEMICAL ENGINEERING JOURNAL
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2024
·
IF 13.3
Haiyang Wang,Hao Zhuang,Chunyan Wu,Bangting Wang,Shijie Zhang,Yao Wang,Li Liu,Lianzhen Yu,Lei Zhang,Su Jing,Zhining Fan
The limited treatment options for colorectal cancer (CRC) and its adverse effects of chemotherapy underscore the urgent need for innovative therapeutic strategies. Focusing on early cellular glycosylation, particularly through the targeting of O-GlcNAcylation with small-molecule inhibitors, emerges as a promising direction. In this study, we developed artificial cell-like nanovesicles with a bilayer membrane structure to encapsulate OSMI-1, a specific inhibitor of O-GlcNAc transferase, aimed at inhibiting CRC progression. To enhance the effectiveness and specificity of the treatment, these self-targeted nanovesicles (T–Se–Lip–OSMI) were reinforced using internal diselenide bridges enabling redox-responsive release within the tumor microenvironment. In vitro assays revealed that T–Se–Lip–OSMI substantially diminished CRC cell proliferation, migration, and invasion. This was further validated using patient-derived organoid models, which highlighted the superior therapeutic efficacy of T–Se–Lip–OSMI over nanovesicles without the diselenide bridge. Subsequent in vivo studies confirmed its targeted delivery and pronounced suppression of tumor growth. Further mechanistic analysis illustrated that the therapeutic effect was mediated through the inhibition of O-GlcNAcylation of the Yes-associated protein (YAP), ultimately preventing the nuclear translocation of YAP and the activation of oncogenic downstream targets. This study leverages the unique properties of the diselenide bridge to enhance stability and functionality of the nanovesicles, offering a novel avenue for targeted CRC treatment.
研究领域:
癌症生物学
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药学
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分子生物学
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纳米技术
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细胞生物学
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高影响因子
ENVIRONMENTAL POLLUTION
·
2025
·
IF 7.3
Ziyuan Li,Pu Chen,Yongfeng Lin,Jingxu Zhang,Jian Ding,Rifat Zubair Ahmed,Xiaoting Jin,Yuxin Zheng
Tris (2-chloroethyl) phosphate (TCEP), recognized as an emerging pollutant, has been frequently detected in human blood. Maintenance of blood homeostasis is indispensable for regulating various physiological states and overall health, yet hematological toxicology of TCEP has not been extensively investigated. Platelets, a vital component of blood, are fundamental in the processes of hemostasis and thrombosis through their activation; thus, this study was designed to elucidate the effects and underlying mechanisms of TCEP on platelet activation. Utilizing an in vivo model, we conducted a proteomic analysis of platelets and found that TCEP exposure inhibited platelet activation. An ex vivo platelet evaluation system was employed to further dissect the processes of platelet activation, revealing that TCEP predominantly suppressed platelet aggregation, degranulation and clot retraction. These processes were highly dependent on energy metabolism, and TCEP was found to decrease ATP levels, primarily by impairing glycolysis and pentose phosphate pathways. Subsequent investigation into the molecular mechanisms revealed that TCEP decreased the activity of phosphofructokinase platelet (PFKP) by enhancing O-linked N-acetylglucosamine (O-GlcNAc) transferase interaction with PFKP. This study is the first to uncover the disruptive effects of TCEP on platelet activation process, providing valuable insights into the assessment of hematologic health risks associated with TCEP-like emerging pollutants exposure.
研究领域:
血液学
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毒理学
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分子生物学
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生物化学
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高影响因子
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