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Characteristic glycopeptides associated with extreme human longevity identified through plasma glycoproteomics

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS 2018年第6期1862卷 1462-1471页

作者： Miura, Yuri Hashii, Noritaka Ohta, Yuki Itakura, Yoko Tsumoto, Hiroki Suzuki, Junya Takakura, Daisuke Abe, Yukiko Arai, Yasumichi Toyoda, Masashi Kawasaki, Nana Hirose, Nobuyoshi Endo, Tamao Tokyo Metropolitan Inst Gerontol Res Team Mech Aging Itabashi Ku 35-2 Sakae Cho Tokyo 1730015 Japan Natl Inst Hlth Sci Div Biol Chem & Biol Kawasaki Ku 3-25-26 Tono Machi Kawasaki Kanagawa 2109501 Japan Tokyo Metropolitan Inst Gerontol Res Team Geriatr Med Itabashi Ku 35-2 Sakae Cho Tokyo 1730015 Japan Keio Univ Sch Med Ctr Supercentenarian Med Res Shinjuku Ku 35 Shinano Machi Tokyo 1608582 Japan Yokohama City Univ Grad Sch Med Life Sci Kanazawa Ku 3-9 Hukuura Yokohama Kanagawa 2360004 Japan Keio Univ Sch Med Ctr Integrated Med Res Shinjuku Ku 35 Shinano Machi Tokyo 1608582 Japan

Background: Glycosylation is highly susceptible to changes of the physiological conditions, and accordingly, is a potential biomarker associated with several diseases and/or longevity. Semi-supercentenarians (SSCs;older than 105 years) are thought to be a model of human longevity. Thus, we performed glycoproteomics using plasma samples of SSCs, and identified proteins and conjugated N-glycans that are characteristic of extreme human longevity. Methods: Plasma proteins from Japanese semi-supercentenarians (SSCs, 106-109 years), aged controls (70-88 years), and young controls (20-38 years) were analysed by using lectin microarrays and liquid chromatography/mass spectrometry (LC/MS). Peak area ratios of glycopeptides to corresponding normalising peptides were subjected to orthogonal projections to latent structures discriminant analysis (OPLS-DA). Furthermore, plasma levels of clinical biomarkers were measured. Results: We found two lectins such as Phaseolus vulgaris, and Erythrina cristagalli (ECA), of which protein binding were characteristically increased in SSCs. Peak area ratios of ECA-enriched glycopeptides were successfully discriminated between SSCs and controls using OPLS-DA, and indicated that tri-antennary and sialylated N-glycans of haptoglobin at Asn207 and Asn211 sites were characterized in SSCs. Sialylated glycans of haptoglobin are a potential biomarker of several diseases, such as hepatocellular carcinoma, liver cirrhosis, and IgA-nephritis. However, the SSCs analysed here did not suffer from these diseases. Conclusions: Tri-antennary and sialylated N-glycans on haptoglobin at the Asn207 and Asn211 sites were abundant in SSCs and characteristic of extreme human longevity. General significance: We found abundant glycans in SSCs, which may be associated with human longevity.

关键词： N-glycosylation lectin microarray LC/MS Longevity Semi-supercentenarian

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Glycomic Analysis Reveals a Conserved Response to Bacterial Sepsis Induced by Different Bacterial Pathogens

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ACS INFECTIOUS DISEASES 2022年第5期8卷 1075-1085页

作者： Heindel, Daniel W. Chen, Shuhui Aziz, Peter, V Chung, Jonathan Y. Marth, Jamey D. Mahal, Lara K. NYU Biomed Res Inst Dept Chem New York NY 10003 USA SBP Med Discovery Inst La Jolla CA 92037 USA Univ Alberta Dept Chem Edmonton AB T6G 2G2 Canada

Sepsis is an extreme inflammatory response to infection that occurs in the bloodstream and causes damage throughout the body. Glycosylation is known to play a role in immunity and inflammation, but the role of glycans in sepsis is not well-defined. Herein, we profiled the serum glycomes of experimental mouse sepsis models to identify changes induced by 4 different clinical bacterial pathogens (Gram-positive: Streptococcus pneumoniae and Staphylococcus aureus, Gramnegative: Escherichia coli and Salmonella Typhimurium) using our lectin microarray technology. We observed global shifts in the blood sera glycome that were conserved across all four species, regardless of whether they were Gram positive or negative. Bisecting GlcNAc was decreased upon sepsis and a strong increase in core 1/3 O-glycans was observed. lectin blot analysis revealed a high molecular weight protein induced in sepsis by all four bacteria as the major cause of the core 1/3 O-glycan shift. Analysis of this band by mass spectrometry identified interalpha-trypsin inhibitor heavy chains (ITIHs) and fibronectin, both of which are associated with human sepsis. Shifts in the glycosylation of these proteins were observed. Overall, our work points toward a common mechanism for bacterially induced sepsis, marked by conserved changes in the glycome.

关键词： serum glycomics core 1 3 O-glycans lectin microarray sepsis interalpha-trypsin inhibitor heavy chains (ITIHs) fibronectin

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XBP1s activation can globally remodel N-glycan structure distribution patterns

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2018年第43期115卷 E10089-E10098页

作者： Wong, Madeline Y. Chen, Kenny Antonopoulos, Aristotelis Kasper, Brian T. Dewal, Mahender B. Taylor, Rebecca J. Whittaker, Charles A. Hein, Pyae P. Dell, Anne Genereux, Joseph C. Haslam, Stuart M. Mahal, Lara K. Shoulders, Matthew D. MIT Dept Chem Cambridge MA 02139 USA Imperial Coll London Dept Life Sci London SW7 2AZ England NYU Dept Chem Biomed Chem Inst New York NY 10003 USA MIT Barbara K Ostrom Bioinformat & Comp Facil 1978 77 Massachusetts Ave Cambridge MA 02139 USA Univ Calif Riverside Dept Chem Riverside CA 92521 USA Expans Technol Inc Cambridge MA 02139 USA Harvard Univ Dept Chem & Chem Biol Cambridge MA 02138 USA Takeda Pharmaceut Inc Mol Biol Cambridge MA 02139 USA

Classically, the unfolded protein response (UPR) safeguards secretory pathway proteostasis. The most ancient arm of the UPR, the IRE1-activated spliced X-box binding protein 1 (XBP1s)-mediated response, has roles in secretory pathway maturation beyond resolving proteostatic stress. Understanding the consequences of XBP1s activation for cellular processes is critical for elucidating mechanistic connections between XBP1s and development, immunity, and disease. Here, we show that a key functional output of XBP1s activation is a cell type-dependent shift in the distribution of N-glycan structures on endogenous membrane and secreted proteomes. For example, XBP1s activity decreased levels of sialylation and bisecting GIcNAc in the HEK293 membrane proteome and secretome, while substantially increasing the population of oligomannose N-glycans only in the secretome. In HeLa cell membranes, stress-independent XBP1s activation increased the population of high-mannose and tetraantennary N-glycans, and also enhanced core fucosylation. mRNA profiling experiments suggest that XBP1s-mediated remodeling of the N-glycome is, at least in part, a consequence of coordinated transcriptional resculpting of N-glycan maturation pathways by XBP1s. The discovery of XBP1s-induced N-glycan structural remodeling on a glycome-wide scale suggests that XBP1s can act as a master regulator of N-glycan maturation. Moreover, because the sugars on cell-surface proteins or on proteins secreted from an XBP1s-activated cell can be molecularly distinct from those of an unactivated cell, these findings reveal a potential new mechanism for translating intracellular stress signaling into altered interactions with the extracellular environment.

关键词： N-glycosylation proteostasis glycoproteome lectin microarray endoplasmic reticulum

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Spatiotemporal changes of tissue glycans depending on localization in cardiac aging

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REGENERATIVE THERAPY 2023年 22卷 68-78页

作者： Itakura, Yoko Hasegawa, Yasuko Kikkawa, Yurika Murakami, Yuina Sugiura, Kosuke Nagai-Okatani, Chiaki Sasaki, Norihiko Umemura, Mariko Takahashi, Yuji Kimura, Tohru Kuno, Atsushi Ishiwata, Toshiyuki Toyoda, Masashi Tokyo Metropolitan Inst Gerontol Res Team Geriatr Med Vasc Med 35-2 Sakae choItabashi ku Tokyo 1730015 Japan Tokyo Metropolitan Inst Gerontol Div Aging & Carcinogenesis Res Team Geriatr Pathol 35-2 Sakae choItabashi ku Tokyo 1730015 Japan Kitasato Univ Sch Sci Dept Biosci Lab Stem Cell Biol 1-15-1 Kitasato Minami ku Sagamihara Kanagawa 2520373 Japan Tokyo Univ Pharm & Life Sci Sch Life Sci Lab Environm Mol Physiol 1432-1 Horinouchi Hachioji Tokyo 1920392 Japan Natl Inst Adv Ind Sci & Technol Cellular & Mol Biotechnol Res Inst 5 Cent Tsukuba 1-1-1 Higashi Tsukuba Ibaraki 3058565 Japan

Introduction: Heart failure is caused by various factors, making the underlying pathogenic mechanisms difficult to identify. Since cardiovascular disease tends to worsen over time, early diagnosis is key for treatment. In addition, understanding the qualitative changes in the heart associated with aging, where information on the direct influences of aging on cardiovascular disease is limited, would also be useful for treatment and diagnosis. To fill these research gaps, the focus of our study was to detect the structural and functional molecular changes associated with the heart over time, with a focus on glycans, which reflect the type and state of ***: We investigated glycan localization in the cardiac tissue of normal mice and their alterations during aging, using evanescent-field fluorescence-assisted lectin microarray, a technique based on lectin-glycan interaction, and lectin ***: The glycan profiles in the left ventricle showed differences between the luminal side (medial) and wall side (lateral) regions. The medial region was characterized by the presence of sialic acid residues. Moreover, agerelated changes in glycan profiles were observed at a younger age in the medial region. The difference in the age-related decrease in the level of a-galactose stained with Griffonia simplicifolia lectin-IB4 in different regions of the left ventricle suggests spatiotemporal changes in the number of microvessels. Conclusions: The glycan profile, which retains diverse glycan structures, is supported by many cell populations, and maintains cardiac function. With further research, glycan localization and changes have the potential to be developed as a marker of the signs of heart failure. (c) 2022, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://***/licenses/by-nc-nd/ 4.0/).

关键词： Cardiac tissue lectin microarray Aging Glycan profile Microvessels

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microarrays in infection and immunity

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CURRENT OPINION IN CHEMICAL BIOLOGY 2007年第3期11卷 306-315页

作者： Maynard, Jennifer A. Myhre, Ryan Roy, Benjamin Univ Minnesota Dept Chem Engn & Mat Sci Minneapolis MN 55455 USA Univ Texas Dept Chem Engn Austin TX 78712 USA

Over the past decade, microarrays have revolutionized the scientific world as dramatically as the internet has changed everyday life. From the initial applications of DNA microarrays to uncover gene expression patterns that are diagnostic and prognostic of cancer, understanding the interplay between immune responses and disease has been a prime application of this technology. More recent efforts have moved beyond genetic analysis to functional analysis of the molecules involved, including identification of immunodominant antigens and peptides as well as the role of post-translational glycosylation. Here, we focus on recent applications of microarray technology in understanding the detailed chemical biology of immune responses to disease in an effort to guide development of vaccines and other protective therapies.

关键词： ACUTE RESPIRATORY SYNDROME RESEQUENCING DNA microarrayS SEROGROUP-B MENINGOCOCCUS GENE-EXPRESSION PROGRAM T-CELL RESPONSES PROTEIN microarrayS OLIGONUCLEOTIDE microarrayS ANTIBODY-RESPONSES lectin microarray GLYCAN ARRAY Immunodominant Antigens Immune Response Immunodominant Epitopes microarray Protein microarrays DNA microarrays maladjustment microarray technology antibody response Functional Analysis Oligonucleotide Arrays Respiratory Syndrome, Acute, Severe Infection everyday life Immunity

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Effectiveness of Selenium on Chondrocyte Glycoprotein Glycosylation Which Play Important Roles in the Pathogenesis of an Endemic Osteoarthritis, Kashin-Beck Disease

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BIOLOGICAL TRACE ELEMENT RESEARCH 2022年第4期200卷 1531-1537页

作者： Wang, Sen Geng, Lingling Zhao, Guanghui Meng, Peilin Yuan, Linlin Guo, Xiong Xi An Jiao Tong Univ Sch Publ Hlth Hlth Sci Ctr Key Lab Trace Elements & Endem DisNatl Hlth Comm Xian Shaanxi Peoples R China Xian Childrens Hosp Dept Rheumatol & Immunol Xian Shaanxi Peoples R China Xi An Jiao Tong Univ Xian Honghui Hosp Hlth Sci Ctr Xian Shaanxi Peoples R China

In this study, we aimed to explore the effectiveness of selenium on the chondrocyte glycoprotein glycosylation which plays important roles in the pathogenesis of Kashin-Beck disease (KBD). Cartilage samples were collected from KBD patients after total knee replacement surgery. Chondrocytes were cultured with sodium selenium. The group of chondrocytes which were cultured without adding sodium selenium was considered as control group. lectin microarray was used to screen the differences in lectin levels between KBD and KBD with selenium groups. Stronger signals for Bandeiraea simplicifolia (BS-I), Hippeastrum hybrid lectin (HHL), Pisum sativum agglutinin (PSA), Psophocarpus tetragonolobus lectin I (PTL-I), Psophocarpus tetragonolobus lectin II (PTL-II), Sophora japonica agglutinin (SJA), Lotus tetragonolobus lectin (LTL), and Triticum vulgaris (WGA) were observed in the KBD group. Meanwhile, Aleuria aurantia lectin (AAL), Lens culinaris agglutinin (LCA), Lycopersicon esculentum (tomato) lectin (LEL), Peanut agglutinin (PNA), and Sambucus nigra lectin (SNA) signals were lower in the KBD group. Selenium may have the function of influence the expression levels of carbohydrate chains Gal alpha 1,3-Gal, high mannose, and GlcNAc.

关键词： Kashin-Beck disease Chondrocyte Selenium Carbohydrate chain lectin microarray

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Development of an all-in-one technology for glycan profiling targeting formalin-embedded tissue sections

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2008年第2期370卷 259-263页

作者： Matsuda, Atsushi Kuno, Atsushi Ishida, Hiroyasu Kawamoto, Toru Shoda, Jun-ichi Hirabayashi, Jun AIST Natl Inst Adv Ind Sci & Technol RCMG Lectin Applicat & Anal Team Tsukuba Ibaraki 3058568 Japan Univ Tsukuba Grad Sch Comprehens Human Sci Dept Gastroenterol Tsukuba Ibaraki 3058577 Japan Mito Med Ctr Ibaraki 3113193 Japan Sojinkai Hosp Ibaraki 3001546 Japan

An ultra-sensitive method for glycan analysis targeting small tissue sections (1.5 mm in diameter) is described as an application of a recently-established lectin microarray technology. The developed system achieved a high level of detection of a tissue section consisting of approximately 500 cells for differential profiling, where both N- and O-glycans attached to a pool of glycoproteins are subjected to multiplex analysis with 43 lectins. By using an optimized protocol for differential glycan analysis, sections of adenocarcinoma (n = 28) and normal epithelia (n = 12) of the colon were analyzed in an all-in-one manner. As a result, Wisteria floribunda agglutinin (WFA) was found to clearly differentiate cancerous from normal epithelia with P < 0.0001. The obtained results correlated well with the subsequent histochemical study using biotinylated WFA. Thus, the developed technology proved to be valid for expanding the lectin microarray applications to tissue-based glycomics, and hence, should accelerate a discovery phase of glycan-related biomarkers. (C) 2008 Elsevier Inc. All rights reserved.

关键词： glycan profiling biomarker lectin microarray tissue array colon cancer formalin-embedded

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Concept, strategy and realization of lectin-based glycan profiling

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JOURNAL OF BIOCHEMISTRY 2008年第2期144卷 139-147页

作者： Hirabayashi, Jun Natl Inst Adv Ind Sci & Technol Res Ctr Med Glycosci Tsukuba Ibaraki 3058568 Japan

lectins are a diverse group of carbohydrate-binding proteins. Each lectin has its own specificity profile. It is believed that lectins exist in all living organisms that produce glycans. From a practical viewpoint, lectins have been used extensively in biochemical fields including proteomics due to their usefulness as detection and enrichment tools for specific glycans. Nevertheless, they have often been underestimated as probes, especially compared with antibodies, because of their low affinity and broad specificity. However, together with the concept of glycomics, such properties of lectins are now considered to be suitable for the task of profiling in order to cover a wider range of ligands. Recently there has been rapid movement in the field of proteomics aimed at the investigation of glycan-related biomarkers. This is partly because of limitations of the present approach of simply following changes in protein-level expression, without paying sufficient attention to the fact and effects of glycosylation. The trend is reflected in the frequent use of lectins in the contexts of glycoprotein enrichment and glycan profiling. However, there are many aspects to be considered in using lectins, which differ considerably from antibodies. In this article, the author, as a developer of two unique methodologies, frontal affinity chromatography (FAC) and the lectin microarray, describes critical points concerning the use of lectins, together with the concept, strategy and means to achieve advances in these emerging glycan profiling technologies.

关键词： biomarker chromatography frontal affinity glycan profiling glycomics lectin microarray

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Functional protein microarray： an ideal platform for investigating protein binding property

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Frontiers in Biology 2012年第4期7卷 336-349页

作者： Shu-Min ZHOU Shanghai Center for Systems Biomedicine Key Laboratory of Systems Biomedicine (Ministry of Education) Shanghai Jiao Tong University Shanghai 200240 China StateKey Laboratory of Oncogenes and Related Genes Shanghai 200240 China Department of Pharmacology and Molecular Sciences Johns Hopkins University School of Medicine Baltimore MD 21205 USA The High-Throughput Biology Center Johns Hopkins University School of Medicine Baltimore MD 21205 USA

Functional protein microarray is an important tool for high-throughput and large-scale systems biology studies. Besides the progresses that have been made for protein microarray fabrication, significant advancements have also been achieved for applying protein microarrays on determining a variety of protein biochemical activities. Among these applications, detection of protein binding properties, such as protein-protein interactions （PPIs）, protein-DNA interactions （PDIs）, protein-RNA interactions, and antigen-antibody interactions, are straightforward and have substantial impacts on many research fields. In this review, we will focus on the recent progresses in protein-protein, protein-DNA, protein-RNA, protein-small molecule, protein-lipid, protein-glycan, and antigen-antibody interactions. We will also discuss the challenges and future directions of protein microarray technologies. We strongly believe that protein microarrays will soon become an indispensible tool for both basic research and clinical applications.

关键词： lectin microarray protein microarray protein-cell interaction protein-DNA interaction （PDI） protein-proteininteraction （PPI）

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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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