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dc.creatorLopandic, Zorana
dc.creatorDragacevic, Luka
dc.creatorPopovic, Dragan
dc.creatorAndjelkovic, Uros
dc.creatorMinic, Rajna
dc.creatorGavrovic-Jankulovic, Marija
dc.date.accessioned2021-11-05T15:10:44Z
dc.date.available2021-11-05T15:10:44Z
dc.date.issued2021
dc.identifier.issn2218-273X
dc.identifier.urihttp://intor.torlakinstitut.com/handle/123456789/615
dc.description.abstractFluorescently 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 microorganismssr
dc.language.isoensr
dc.publisherMDPIsr
dc.relationinfo:eu-repo/grantAgreement/MESTD/inst-2020/200168/RS//sr
dc.relationinfo:eu-repo/grantAgreement/MESTD/inst-2020/200177/RS//sr
dc.relationinfo:eu-repo/grantAgreement/MESTD/inst-2020/200026/RS//sr
dc.rightsopenAccesssr
dc.sourceBiomoleculessr
dc.subjectbanana lectinsr
dc.subjecteGFPsr
dc.subjectfluorescencesr
dc.subjectviral glycoproteinssr
dc.subjectinfluenza vaccinesr
dc.subjectflorescence-linked lectin sorbent assaysr
dc.subjectSalmonella strainssr
dc.titleBanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganismssr
dc.typearticlesr
dc.rights.licenseBYsr
dc.citation.issue2
dc.citation.spage180
dc.citation.volume11(2)
dc.citation.volume11
dc.identifier.doi10.3390/biom11020180
dc.identifier.scopus2-s2.0-85099852243
dc.identifier.wos000622122600001
dc.type.versionpublishedVersionsr


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