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BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms
dc.creator | Lopandić, Zorana | |
dc.creator | Dragačević, Luka | |
dc.creator | Popović, Dragan M. | |
dc.creator | Anđelković, Uroš | |
dc.creator | Minić, Rajna | |
dc.creator | Gavrović-Jankulović, Marija | |
dc.date.accessioned | 2021-05-17T13:04:04Z | |
dc.date.available | 2021-05-17T13:04:04Z | |
dc.date.issued | 2021 | |
dc.identifier.issn | 2218-273X | |
dc.identifier.uri | https://cer.ihtm.bg.ac.rs/handle/123456789/4629 | |
dc.description.abstract | Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms. | sr |
dc.language.iso | en | sr |
dc.publisher | MDPI | sr |
dc.relation | The bilateral scientific project financed by Ministries of Sciences of Croatia and Serbia | sr |
dc.relation | info:eu-repo/grantAgreement/MESTD/inst-2020/200168/RS// | sr |
dc.relation | info:eu-repo/grantAgreement/MESTD/inst-2020/200177/RS// | sr |
dc.relation | info:eu-repo/grantAgreement/MESTD/inst-2020/200026/RS// | sr |
dc.rights | openAccess | sr |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
dc.source | Biomolecules | sr |
dc.subject | Banana lectin | sr |
dc.subject | EGFP | sr |
dc.subject | Florescence-linked lectin sorbent assay | sr |
dc.subject | Fluorescence | sr |
dc.subject | Influenza vaccine | sr |
dc.subject | Salmonella strains | sr |
dc.subject | Viral glycoproteins | sr |
dc.subject | enhanced green fluorescent protein | sr |
dc.subject | glycoprotein | sr |
dc.subject | hemagglutin | sr |
dc.subject | influenza vaccine | sr |
dc.subject | amino acid sequence | sr |
dc.subject | antigen binding | sr |
dc.subject | Article | sr |
dc.subject | binding affinity | sr |
dc.subject | binding site | sr |
dc.subject | biotinylation | sr |
dc.subject | circular dichroism | sr |
dc.subject | controlled study | sr |
dc.subject | crystal structure | sr |
dc.subject | enzymatic assay | sr |
dc.subject | epitope mapping | sr |
dc.subject | Escherichia coli | sr |
dc.subject | low cytometry | sr |
dc.subject | fluorescence linked lectin sorbent assay | sr |
dc.subject | glycosylation | sr |
dc.subject | human | sr |
dc.subject | human cell | sr |
dc.subject | IC50 | sr |
dc.subject | immunoblotting | sr |
dc.subject | in vitro study | sr |
dc.subject | ion exchange chromatography | sr |
dc.subject | lectin binding | sr |
dc.subject | mass spectrometry | sr |
dc.subject | molecular mechanics | sr |
dc.subject | molecular model | sr |
dc.subject | nucleotide sequence | sr |
dc.subject | protein protein interaction | sr |
dc.subject | protein purification | sr |
dc.subject | protein structure | sr |
dc.subject | Salmonella enterica serovar Typhi | sr |
dc.subject | structure activity relation | sr |
dc.subject | structure analysis | sr |
dc.subject | surface plasmon resonance | sr |
dc.subject | X ray crystallography | sr |
dc.title | BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms | sr |
dc.type | article | sr |
dc.rights.license | BY | sr |
dcterms.abstract | Гавровић-Јанкуловић, Марија; Минић, Рајна; Aнђелковић, Урош; Поповић, Драган М.; Драгачевић, Лука; Лопандић, Зорана; | |
dc.citation.volume | 11 | |
dc.citation.issue | 2 | |
dc.citation.spage | 180 | |
dc.citation.rank | M22~ | |
dc.identifier.pmid | 33525574 | |
dc.identifier.doi | 10.3390/biom11020180 | |
dc.identifier.fulltext | https://cer.ihtm.bg.ac.rs/bitstream/id/20404/biomolecules-11-00180.pdf | |
dc.identifier.scopus | 2-s2.0-85099852243 | |
dc.identifier.wos | 000622122600001 | |
dc.type.version | publishedVersion | sr |