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Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray

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CARBOHYDRATE RESEARCH 2014年第1期389卷 123-133页

作者： Kilcoyne, Michelle Twomey, Marcus E. Gerlach, Jared Q. Kane, Marian Moran, Anthony P. Joshi, Lokesh Natl Univ Ireland Natl Ctr Biomed Engn Sci Glycosci Grp Galway Ireland Natl Univ Ireland Sch Nat Sci Galway Ireland

Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barre syndrome (GBS). While C. jejuni is pathogenic in humans, it is a commensal in avian hosts. Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. C. jejuni strains 81116 and 81-176 were cultured at 37 and 42 degrees C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37 degrees C. The profiles of both strains varied and were distinguishable at both temperatures. At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures w

关键词： Campylobacter jejuni lectin microarray Virulence factors Lipooligosaccharide Lipopolysaccharide Capsular polysaccharide

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Altered N-Glycan Expression Profile in Epithelial-to-Mesenchymal Transition of NMuMG Cells Revealed by an Integrated Strategy Using Mass Spectrometry and Glycogene and lectin microarray Analysis

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JOURNAL OF PROTEOME RESEARCH 2014年第6期13卷 2783-2795页

作者： Tan, Zengqi Lu, Wei Li, Xiang Yang, Ganglong Guo, Jia Yu, Hanjie Li, Zheng Guan, Feng Jiangnan Univ Sch Biotechnol Minist Educ Key Lab Carbohydrate Chem & Biotechnol Wuxi 214122 Jiangsu Peoples R China Jiangnan Univ Wuxi Med Sch Wuxi 214122 Jiangsu Peoples R China NW Univ Xian Coll Life Sci Lab Funct Glyc Xian 710069 Peoples R China

Epithelial-to-mesenchymal transition (EMT) is an essential biological process that occurs in embryonic development, metastatic diseases, and cancer progression. Altered expression of glycans is known to be associated with cancer progression. No studies to date have presented global analysis of the precise variation of N-glycans in EMT. We describe here the profile of N-glycans and glycogene expression in the EMT process induced by transforming growth factor-beta 1 (TGF beta 1) in a normal mouse mammary gland epithelial (NMuMG) cell model. An integrated strategy with a combination of mass spectrometry, glycogene microarray analysis, and lectin microarray analysis was applied, and results were confirmed by lectin histochemistry and quantitative real-time PCR. In TGF beta-induced EMT, levels of high-mannose-type N-glycans were enhanced, antennary N-glycans, and fucosylation were suppressed, and bisecting GIcNAc N-glycans were greatly suppressed. The expression of seven N-glycan-related genes was significantly changed. The products of glycogenes ALG9, MGAT3, and MGAT4B appeared to contribute to the observed alteration of N-glycans. The findings indicate that dysregulation of N-glycan synthesis plays a role in the EMT process. Systematic glycomic analysis based on the combination of techniques described here is expected to facilitate the discovery of the aberrant N-glycosylation in tumor progression and provide essential information in systems glycobiology.

关键词： N-glycan glycogene mass spectrometry gene microarray lectin microarray high throughput

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Comprehensive profiling of accessible surface glycans of mammalian sperm using a lectin microarray

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CLINICAL PROTEOMICS 2014年第1期11卷 10-10页

作者： Xin, Ai-Jie Cheng, Li Diao, Hua Wang, Peng Gu, Yi-Hua Wu, Bin Wu, Yan-Cheng Chen, Guo-Wu Zhou, Shu-Min Guo, Shu-Juan Shi, Hui-Juan Tao, Sheng-Ce Shanghai Jiao Tong Univ Minist Educ Key Lab Syst Biomed Shanghai Ctr Syst Biomed Shanghai 200240 Peoples R China State Key Lab Oncogenes & Related Genes Shanghai 200240 Peoples R China Shanghai Jiao Tong Univ Sch Biomed Engn Shanghai 200240 Peoples R China Fudan Univ Obstet & Gynecol Hosp Shanghai Key Lab Female Reprod Endocrine Related Shanghai Jiai Genet & IVF Inst Shanghai 200011 Peoples R China SIPPR China Natl Populat & Family Planning Key Lab Cont Shanghai 200032 Peoples R China E China Univ Sci & Technol State Key Lab Bioreactor Engn Shanghai 20037 Peoples R China

It is well known that cell surface glycans or glycocalyx play important roles in sperm motility, maturation and fertilization. A comprehensive profile of the sperm surface glycans will greatly facilitate both basic research (sperm glycobiology) and clinical studies, such as diagnostics of infertility. As a group of natural glycan binders, lectin is an ideal tool for cell surface glycan profiling. However, because of the lack of effective technology, only a few lectins have been tested for lectin-sperm binding profiles. To address this challenge, we have developed a procedure for high-throughput probing of mammalian sperm with 91 lectins on lectin microarrays. Normal sperm from human, boar, bull, goat and rabbit were collected and analyzed on the lectin microarrays. Positive bindings of a set of similar to 50 lectins were observed for all the sperm of 5 species, which indicated a wide range of glycans are on the surface of mammalian sperm. Species specific lectin bindings were also observed. Clustering analysis revealed that the distances of the five species according to the lectin binding profiles are consistent with that of the genome sequence based phylogenetic tree except for rabbit. The procedure that we established in this study could be generally applicable for sperm from other species or defect sperm from the same species. We believe the lectin binding profiles of the mammalian sperm that we established in this study are valuable for both basic research and clinical studies.

关键词： Sperm Glycocalyx Surface glycan lectin microarray

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lectins: Versatile proteins in plant life with diverse applications in immunomodulation, antibacterial defense, and disease-fighting

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JOURNAL OF CARBOHYDRATE CHEMISTRY 2024年第7-9期43卷 243-288页

作者： Jeyachandran, Sivakamavalli Vibhute, Prachi Saveetha Univ Saveetha Dent Coll & Hosp Saveetha Inst Med & Tech Sci SIMATS Lab Biotechnol & Biosignal TransductDept Orthodon Chennai Tamil Nadu India Natl Coll Autonomous PG & Res Dept Biotechnol & Microbiol Tiruchirappalli Tamil Nadu India

lectins, diverse proteins adept at detecting and binding carbohydrates without altering them, are pivotal in plant life. They facilitate cell communication, development, and defense mechanisms. This article explores lectins' varied biological roles, focusing on their versatility and significance. Plant lectins exhibit diverse biochemical traits, impacting immune regulation and possessing antibacterial properties vital in plant defense. Additionally, they showcase disease-fighting potential. Their multifaceted nature, spanning biochemical properties, and functional roles underscores their importance in intricate biological processes. This concise review highlights wide-ranging applications of plant lectins, emphasizing their crucial role in immunomodulation, antibacterial functions, and contributions to plant health and defense mechanisms.

关键词： Antimicrobial activity anti-tumor activity carbohydrate specific binding lectin microarray plant lectins

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Glycopattern comparison of sperm and exosomes from conventional in vitro fertilization and rescue intracytoplasmic sperm injection: Enhancing sperm motility through exosome-mediated glycan modulation

VIEW

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VIEW 2025年

作者： Ramzan, Muhammad Anas Yang, Zhedong Cao, Li Zhang, Jinyuan Wu, Fei Xin, Miaomiao Guo, Bo Wang, Lumin Zhao, Lingyu Qin, Yannan Liu, Chengcheng Xi An Jiao Tong Univ Inst Genet & Dev Biol Translat Med Inst Xian 710301 Peoples R China Northwest Womens & Childrens Hosp Reprod Med Ctr Xian Peoples R China Xi An Jiao Tong Univ Affiliated Hosp 2 Dept Oncol Xian Peoples R China Xi An Jiao Tong Univ Inst Infect & Immun Translat Med Inst Xian 710301 Peoples R China

Infertility is a global issue, with male factors contributing to approximately 50% of cases. While assisted reproductive technologies (ART) resolve 30%-50% of cases, 2%-5% of IVF failures require rescue intracytoplasmic sperm injection (RICSI). Sperm glycocalyx is critical for capacitation and fertilization, and sperm-derived exosomes, essential for cell communication, remain underexplored. This study used lectin microarrays and fluorescence lectin histochemistry to analyze glycopatterns in sperm and exosomes from IVF and RICSI groups, and assessed RICSI sperm motility and glycan expression after exposure to IVF-derived exosomes. Clinical data from 2023 at Northwest Women and Children's Hospital, China, showed that RICSI couples had longer infertility durations, and younger ages, required lower gonadotropin doses, and retrieved more oocytes, despite lower sperm quality. However, pregnancy rates were similar between RICSI and IVF groups. Glycopattern analysis revealed that alpha GalNAc, Tn antigen, and GalNAc alpha 1-3(Fuc alpha 1-2)Gal (DBA-recognized) glycans were downregulated in RICSI sperm, while Gal beta 1-3GalNAc alpha-Ser/Thr (T), GalNAc alpha-Ser/Thr (Tn), and sialyl-T (Jacalin-recognized) were upregulated in RICSI exosomes compared to IVF. Exosomes showed higher expression of PHA-L-, SBA-, and UEA-I-bound glycans, but lower AAL-bound glycans than sperm. DBA is primarily bound to the neck, acrosomal region, plasma membrane, and tail of IVF sperm, but its binding was significantly reduced in RICSI sperm. Functionally, IVF exosomes enhanced RICSI sperm motility in a dose-dependent manner by upregulating DBA-recognized glycans. These findings provide insights into biomarkers, IVF optimization, and glycosylation's role in sperm and exosome-related infertility, suggesting exosome-based therapy for improving RICSI sperm motility.

关键词： glycopattern in vitro fertilization lectin microarray rescue intracytoplasmic sperm injection sperm sperm-derived exosome

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An improved evanescent fluorescence scanner suitable for high-resolution glycome mapping of formalin-fixed paraffin-embedded tissue sections

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ANALYTICAL AND BIOANALYTICAL CHEMISTRY 2023年第28期415卷 6975-6984页

作者： Boottanun, Patcharaporn Nagai-Okatani, Chiaki Nagai, Misugi Ungkulpasvich, Umbhorn Yamane, Shinjiro Yamada, Masao Kuno, Atsushi Natl Inst Adv Ind Sci & Technol Cellular & Mol Biotechnol Res Inst Mol & Cellular Glycoprote Res Grp Tsukuba Ibaraki 3058565 Japan GlycoTechnica Ltd 101 Hiranobiru3 5-28-6 Utsukushigaoka Aoba Ku Yokohama Kanagawa 2250002 Japan EMUKK LLC 2-21-19 Matsunoki Kuwana Mie 5110902 Japan

lectin microarray (LMA) is a high-throughput platform that enables the rapid and sensitive analysis of N- and O-glycans attached to glycoproteins in biological samples, including formalin-fixed paraffin-embedded (FFPE) tissue sections. Here, we evaluated the sensitivity of the advanced scanner based on the evanescent-field fluorescence principle, which is equipped with a 1x infinity correction optical system and a high-end complementary metal-oxide semiconductor (CMOS) image sensor in digital binning mode. Using various glycoprotein samples, we estimated that the mGSR1200-CMOS scanner has at least fourfold higher sensitivity for the lower limit of linearity range than that of a previous charge-coupled device scanner (mGSR1200). A subsequent sensitivity test using HEK293T cell lysates demonstrated that cell glycomic profiling could be performed with only three cells, which has the potential for the glycomic profiling of cell subpopulations. Thus, we examined its application in tissue glycome mapping, as indicated in the online LM-GlycomeAtlas database. To achieve fine glycome mapping, we refined the laser microdissection-assisted LMA procedure to analyze FFPE tissue sections. In this protocol, it was sufficient to collect 0.1 mm(2) of each of the tissue fragments from 5-mu m-thick sections, which differentiated the glycomic profile between the glomerulus and renal tubules of a normal mouse kidney. In conclusion, the improved LMA enables high-resolution spatial analysis, which expands the possibilities of its application classifying cell subpopulations in clinical FFPE tissue specimens. This will be used in the discovery phase for the development of novel glyco-biomarkers and therapeutic targets, and to expand the range of target diseases.

关键词： lectin microarray Evanescent fluorescence Glycan profiling Endogenous glycoproteins Tissue sections Cell subpopulations

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BMP-1-induced GBA1 nuclear accumulation provokes CCN2 mRNA expression via importin-β-mediated nucleocytoplasmic pathway

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JOURNAL OF CELL COMMUNICATION AND SIGNALING 2023年第2期17卷 263-274页

作者： Muromachi, Koichiro Nakano, Rei Fujita-Yoshigaki, Junko Sugiya, Hiroshi Tani-Ishii, Nobuyuki Kanagawa Dent Univ Grad Sch Dent Dept Endodont Yokosuka Kanagawa 2388580 Japan RIKEN Yokohama Inst Ctr Integrat Med Sci Lab Mucosal Immun Yokohama Kanagawa 2300045 Japan Nihon Univ Coll Bioresource Sci Dept Vet Med Lab Vet Radiol Fujisawa Kanagawa 2520880 Japan Nihon Univ Dept Physiol Sch Dent Matsudo Matsudo Chiba 2718587 Japan Japan Anim Specialty Med Inst JASMINE Yokohama Kanagawa 2240001 Japan

Bone morphogenetic protein (BMP)-1 is expressed by odontoblasts in the dentin-pulp complex. Although the functional effects of BMP-1 on the maturation of various preforms of proteins and enzymes involved in initiating mineralization have been widely observed, how BMP-1 affects cellular molecules remains unknown. We performed a comprehensive analysis of BMP-1-altered glycome profiles and subsequent assays to identify the target glycoproteins in human dental pulp cells (hDPCs) by a glycomic approach. In the presence of BMP-1, a lectin microarray analysis and lectin-probed blotting showed that alpha 2,6-sialylation was significantly attenuated in insoluble fractions from hDPCs. Six proteins were identified by a mass spectrometry analysis of alpha 2,6-sialylated glycoproteins purified using a lectin column. Among them, glucosylceramidase (GBA1) was found to accumulate in the nuclei of hDPCs in the presence of BMP-1. Moreover, BMP-1-induced cellular communication network factor (CCN) 2 expression, which is well known as the osteogenesis/chondrogenesis marker, was significantly suppressed in the cells transfected with GBA1 siRNA. Furthermore, importazole, a potent inhibitor of importin-beta-mediated nuclear import significantly suppressed BMP-1-induced GBA1 nuclear accumulation and BMP-1-induced CCN2 mRNA expression, respectively. Thus, BMP-1 facilitates the accumulation of GBA1 in the nucleus through the reduction of alpha 2,6-sialic acid, which potentially contributes to the transcriptional regulation of the CCN2 gene via importin-beta-mediated nuclear import pathway in hDPCs. Our results offer new insights into the role of the BMP-1-GBA1-CCN2 axis in the development, tissue remodeling, and pathology of dental/craniofacial diseases. [GRAPHICS] .

关键词： Glycomics lectin microarray LC-MS MS analysis Sialylation Glycosphingolipid

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Toward spatial glycomics and glycoproteomics: Innovations and applications

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BBA ADVANCES 2025年 7卷 100146页

作者： Boottanun, Patcharaporn Fuseya, Sayaka Kuno, Atsushi Natl Inst Adv Ind Sci & Technol Cellular & Mol Biotechnol Res Inst Mol & Cellular Glycoprote Res Grp Tsukuba Ibaraki 3058565 Japan Soka Univ Glycan & Life Syst Integrat Ctr GaLSIC Hachioji Tokyo 1928577 Japan

In this mini review, we provide an overview of the challenging field of spatial glycomics/glycoproteomics. Owing to their complexity, sophisticated analytical methods and innovative technologies are needed to advance this field. An agile development approach enables unraveling aberrant glycosylations, glycobiomarkers, and glycotargets for spatial imaging, diagnosis, and therapeutic purposes. We discuss glycopathology and tissue glycomic profiling using highly sensitive lectin-based analyses and introduce deep visual proteomics for glycomic/glycoproteomic sample preparation. Additionally, we highlight the importance of leveraging laser microdissection and artificial intelligence-driven visual software for cell-type assignment and automation techniques, which are crucial for advancing glycomics/glycoproteomics.

关键词： Automated analysis Artificial intelligence Glycomics Glycoproteomics Imaging mass cytometry lectin microarray Spatial omics

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Glycosylation profiles of breast cancer cells may represent clonal variations of multiple organ metastases

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CLINICAL & EXPERIMENTAL METASTASIS 2024年第3期41卷 267-270页

作者： Horimoto, Yoshiya Hlaing, May Thinzar Saeki, Harumi Denda-Nagai, Kaori Ishii-Schrade, Katrin Fujihira, Haruhiko Abe, Masaaki Noji, Miki Shichino, Shigeyuki Saito, Mitsue Irimura, Tatsuro Juntendo Univ Dept Breast Oncol Grad Sch Med Tokyo Japan Juntendo Univ Grad Sch Med Dept Human Pathol Tokyo Japan Tokyo Med Univ Dept Breast Oncol & Surg Tokyo Japan Juntendo Univ Div Glycobiol Grad Sch Med 2-1-1 HongoBunkyo ku Tokyo 1138421 Japan RIKEN Glycometabol Biochem Lab Cluster Pioneering Res CPR Wako Japan Juntendo Univ Grad Sch Med Dept Pathol & Oncol Tokyo Japan Tokyo Univ Sci Res Inst Biomed Sci Div Mol Regulat Inflammatory & Immune Dis Tokyo Japan

Glycosylation changes of cancer cells are known to be associated with malignant progression and metastases and potentially determine the organ-selective nature of metastasis as theorized by Paget (Lancet 1:571-573, 1889). Cellular glycans play a variety of roles in the processes of metastasis and may be unique to the cells that metastasize to different organs. We analyzed the glycosylation profiles of the primary tumor and tumors metastasized to lymph node, liver, lung, brain, bone, thyroid, kidney, adrenal, small intestine and pancreas in an autopsy case of breast cancer employing a lectin microarray with 45 lectins. Clustering analysis of the data revealed that metastatic breast cancer cells were categorized into several clusters according to their glycosylation profiles. Our results provide a biological basis to understand differential phenotypes of metastatic breast cancer cells potentially reflecting clonal origin, which does not directly reflect genomic or genetic changes or microenvironmental effects but connects to glycosylation profiles.

关键词： Breast cancer Glycosylation lectin microarray Multiorgan metastasis Clonal variation

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Sweet Secrets: Exploring Novel Glycans and Glycoconjugates in the Extracellular Polymeric Substances of "Candidatus Accumulibacter"

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ACS ES&T WATER 2024年第8期4卷 3391-3399页

作者： Paez-Watson, Timothy Tomas-Martinez, Sergio de Wit, Roeland Keisham, Sunanda Tateno, Hiroaki van Loosdrecht, Mark C. M. Lin, Yuemei Delft Univ Technol Dept Biotechnol NL-2629 HZ Delft Netherlands Natl Inst Adv Ind Sci & Technol Cellular & Mol Biotechnol Res Inst Tsukuba Ibaraki 3058566 Japan

Biological wastewater treatment relies on microorganisms that grow as flocs, biofilms, or granules for efficient separation of biomass from cleaned water. This biofilm structure emerges from the interactions between microbes that produce, and are embedded in, extracellular polymeric substances (EPS). The true composition and structure of the EPS responsible for dense biofilm formation are still obscure. We conducted a bottom-up approach utilizing advanced glycomic techniques to explore the glycan diversity in the EPS from a highly enriched "Candidatus Accumulibacter" granular sludge. Rare novel sugar monomers such as N-Acetylquinovosamine (QuiNAc) and 2-O-Methylrhamnose (2-OMe-Rha) were identified to be present in the EPS of both enrichments. Further, a high diversity in the glycoprotein structures of said EPS was identified by means of lectin based microarrays. We explored the genetic potential of "Ca. Accumulibacter" high quality metagenome assembled genomes (MAGs) to showcase the shortcoming of top-down bioinformatics based approaches at predicting EPS composition and structure, especially when dealing with glycans and glycoconjugates. This work suggests that more bottom-up research is necessary to understand the composition and complex structure of EPS in biofilms since genome based inference cannot directly predict glycan structures and glycoconjugate diversity.

关键词： glycans glycoproteins glycomics extracellular polymeric substances lectin microarray

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