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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产品关联
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
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
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2020
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IF 5.339
Xing-Hui Ren,Hai-Yan Wang,Si Li,Xi-Wen He,Wen-You Li,Yu-Kui Zhang
Early and accurate detection of breast cancer plays an important role in improving the survival rates of patients. In this work, we designed and synthesized the Gal-NAc-imprinted nanoparticles (GIPs) via boronate-affinity glycan-oriented surface imprinting strategy. Molecularly imprinted polymers (MIPs) were hybridized with fluorescent silicon nanoparticles (SiNPs) to target Tn antigens. However, the single fluorescent imaging mode is not conducive to obtaining accurate diagnosis, due to its poor tissue penetration. To resolve this obstacle, doping gadolinium (Gd) into SiNPs was adopted to emerge an extra significant magnetic resonance (MR) signal, achieving highly sensitive fluorescence imaging and magnetic resonance imaging (MRI) with high spatial resolution. GIPs had uniform particle size around 31.8 nm, and exhibited satisfactory fluorescence stability. The maximum adsorption capacity of GIPs was 1.15 μM/g with a high imprinting factor (IF) of 7.5. Confocal laser scanning microscope imaging revealed that the GIPs had excellent specific recognition ability with a low cytotoxicity. GIPs also showed an outstanding MR performance on cancer cells. Therefore, the synthesized nanoparticles had desirable performance in dual-model imaging to specifically target recognition cancer cells. It may have a tremendous potential in real biological samples.
研究领域:
材料科学
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纳米技术
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癌症研究
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生物医学工程
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医学成像
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
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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
✓ 已验证
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）