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Article

BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms

1
Department of Biochemistry, Faculty of Chemistry, University of Belgrade, 11000 Belgrade, Serbia
2
Institute of Virology, Vaccines and Sera, 11152 Belgrade, Serbia
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Institute of Chemistry, Technology and Metallurgy, National Institute of the Republic of Serbia, University of Belgrade, 11000 Belgrade, Serbia
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Department of Biotechnology, University of Rijeka, 5100 Rijeka, Croatia
*
Author to whom correspondence should be addressed.
Academic Editor: Andrey P. Anisimov
Biomolecules 2021, 11(2), 180; https://doi.org/10.3390/biom11020180
Received: 18 November 2020 / Revised: 21 January 2021 / Accepted: 22 January 2021 / Published: 28 January 2021
(This article belongs to the Section Biological Factors)
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. View Full-Text
Keywords: banana lectin; eGFP; fluorescence; viral glycoproteins; influenza vaccine; florescence-linked lectin sorbent assay; Salmonella strains banana lectin; eGFP; fluorescence; viral glycoproteins; influenza vaccine; florescence-linked lectin sorbent assay; Salmonella strains
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MDPI and ACS Style

Lopandić, Z.; Dragačević, L.; Popović, D.; Andjelković, U.; Minić, R.; Gavrović-Jankulović, M. BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. Biomolecules 2021, 11, 180. https://doi.org/10.3390/biom11020180

AMA Style

Lopandić Z, Dragačević L, Popović D, Andjelković U, Minić R, Gavrović-Jankulović M. BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. Biomolecules. 2021; 11(2):180. https://doi.org/10.3390/biom11020180

Chicago/Turabian Style

Lopandić, Zorana, Luka Dragačević, Dragan Popović, Uros Andjelković, Rajna Minić, and Marija Gavrović-Jankulović. 2021. "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms" Biomolecules 11, no. 2: 180. https://doi.org/10.3390/biom11020180

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