Glycoproteomic analysis of WGA‐bound glycoprotein biomarkers in sera from patients with lung adenocarcinoma

Hongsachart, Piyorot; Liu, Rosa Huang; Sinchaikul, Supachok; Pan, Fu-Ming; Phutrakul, Suree; Chuang, Yu-Min; Yu, Chong‐Jen; Chen, Shui‐Tein

doi:10.1002/elps.200800405

Cited by 45 publications

(75 citation statements)

References 60 publications

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“…Proteins that contain the desired glycans were enriched from cell lysates or a protein mixture and quantitatively analyzed by mass spectrometry. Glycoproteomic approaches have been applied to identify new tumor markers (25,41,42). In this study, we successfully identified 16 EV71-interacting proteins by glycoproteomic approaches.…”

Section: Discussionmentioning

confidence: 99%

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Wang

Huang

et al. 2015

J Virol

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Because the pathogenesis of enterovirus 71 (EV71) remains mostly ambiguous, identifying the factors that mediate viral binding and entry to host cells is indispensable to ultimately uncover the mechanisms that underlie virus infection and pathogenesis. Despite the identification of several receptors/attachment molecules for EV71, the binding, entry, and infection mechanisms of EV71 remain unclear. Herein, we employed glycoproteomic approaches to identify human nucleolin as a novel binding receptor for EV71. Glycoproteins purified by lectin chromatography from the membrane extraction of human cells were treated with sialidase, followed by immunoprecipitation with EV71 particles. Among the 16 proteins identified by tandem mass spectrometry analysis, cell surface nucleolin attracted our attention. We found that EV71 interacted directly with nucleolin via the VP1 capsid protein and that an antinucleolin antibody reduced the binding of EV71 to human cells. In addition, the knockdown of cell surface nucleolin decreased EV71 binding, infection, and production in human cells. Furthermore, the expression of human nucleolin on the cell surface of a mouse cell line increased EV71 binding and conferred EV71 infection and production in the cells. These results strongly indicate that human nucleolin can mediate EV71 binding to and infection of cells. Our findings also demonstrate that the use of glycoproteomic approaches is a reliable methodology to discover novel receptors for pathogens. IMPORTANCEOutbreaks of EV71 have been reported in Asia-Pacific countries and have caused thousands of deaths in young children during the last 2 decades. The discovery of new EV71-interacting molecules to understand the infection mechanism has become an emergent issue. Hence, this study uses glycoproteomic approaches to comprehensively investigate the EV71-interacting glycoproteins. Several EV71-interacting glycoproteins are identified, and the role of cell surface nucleolin in mediating the attachment and entry of EV71 is characterized and validated. Our findings not only indicate a novel target for uncovering the EV71 infection mechanism and anti-EV71 drug discovery but also provide a new strategy for virus receptor identification.

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Section: Discussionmentioning

confidence: 99%

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Wang

Huang

et al. 2015

J Virol

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“…WGA lectin affinity, for example, was used to enrich glycoproteins in serum samples and then gel electrophoresis and MS analysis was used to identify 39 differentially expressed proteins and suggested as possible biomarkers in lung cancer development and progression. WGA was found to capture more glycoproteins from the serum samples than six other lectins that were tested [186]. The immobilization of lectins on monolithic support was mostly done by covalent binding of amines on lysine residues to aldehyde-functionalized surfaces.…”

Section: Lectin Affinitymentioning

confidence: 99%

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

Alla

Stine

2015

Chromatography

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Monolithic column materials offer great advantages as chromatographic media in bioseparations and as solid-supports in biocatalysis. These single-piece porous materials have an interconnected ligament structure that limits the void volume inside the column, thus increasing the efficiency without sacrificing the permeability. The preparation of monolithic materials is easy, reproducible and has available a wide range of chemistries to utilize. Complex, heterogeneous and isobaric glycan structures require preparation methods that may include glycan release, separation and enrichment prior to a comprehensive and site-specific glycosylation analysis. Monolithic column materials aid that demand, as shown by the results reported by the research works presented in this review. These works include selective capture of glycans and glycoproteins via their interactions with lectins, boronic acids, hydrophobic, and hydrophilic/polar functional groups on monolith surfaces. It also includes immobilization of enzymes trypsin and PNGase F on monoliths to digest and deglycosylate glycoproteins and glycopeptides, respectively. The use of monolithic capillary columns for glycan separations through nano-liquid chromatography (nano-LC) and capillary electrochromatography (CEC) and coupling these columns to MS instruments to create multidimensional systems show the potential in the development of miniaturized, high-throughput and automated systems of glycan separation and analysis.

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“…Byly nalezeny tři glykoproteiny ve zvýšené koncentraci (adiponektin, ceruloplasmin a glykofosfatidyl-inositol-80) a dva glykoproteiny ve snížené koncentraci (cyclin H a Fyt) přímo související s vývojem adenokarcinomu plic, které by mohly být užitečné pro včasnou detekci onemocnění nebo sledování jeho progrese [47].…”

Section: Závěrunclassified

New Trends in the Study of Protein Glycosylation in Oncological Diseases

Zahradníková¹,

Hernychová²,

Vojtěšek³

et al. 2014

Klin Onkol

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Regionální centrum aplikované molekulární onkologie, Masarykův onkologický ústav, Brno SouhrnGlykomika a glykoproteomika představují relativně nové směry pro analýzy komplexních biologických vzorků a jejich důležitost neustále roste. Tyto oblasti jsou komplementární k dalším zavedeným přístupům, např. ke genomickému profi lování a proteomice. Glykoproteiny jsou stále více uznávány jako důležité molekuly účastnící se buněčných interakcí a adhezí. Výskyt strukturních změn v glykanových částech se zdá být typický pro různé typy rakoviny. Násle-dující souhrn se zabývá aktuálními trendy v glykomickém profi lování a glykoproteomickém výzkumu bio logických tekutin a tkání se zaměřením na rakovinu. Použité metody jsou založeny na principech kapilárních separačních technik, hmotnostní spektrometrie a glykanových a lektinových čipů. Všechny zmíněné metody mají značný potenciál pro využití v dia gnostických a prediktivních vyšetřeních. Klíčová slovaglykomika -glykopeptidy -rakovina -kapalinová chromatografi e -hmotnostní spektrometrie -kapilární elektroforéza -glykanové profi lování -čipové analýzy SummaryGlycomics and glycoproteomics represent relatively new directions in detail analyses of complex bio logical media. These areas of increasing importance to cancer research complement the more established genomic profi ling and proteomics. Glycoproteins are being increasingly recognized as important in cellular interactions and adhesion. Structural alterations of their glycan moieties seem to occur in diff erent cancer conditions. We review current directions in glycomic profi ling and glycoproteomic investigations of bio logical fl uids and tissues pertaining to cancer. The used methods rely on capillary separation techniques, mass spectrometry, and the glycan and lectin arrays. They all show considerable promise for new dia gnostic and prognostic measurements.

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Glycoproteomic analysis of WGA‐bound glycoprotein biomarkers in sera from patients with lung adenocarcinoma

Cited by 45 publications

References 60 publications

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

New Trends in the Study of Protein Glycosylation in Oncological Diseases

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