Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers
Receptor-mediated transcytosis is an elegant and promising strategy for drug delivery across biological barriers. Here, we describe a novel ligand–receptor pair based on a dimeric, engineered derivative of the <i>Pseudomonas aeruginosa</i> lectin LecA, here termed Di-LecA, and the host c...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/15/1/225 |
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| author | Maokai Xu Maria Antonova Pavel Salavei Katharina Illek Ana Valeria Meléndez Ramin Omidvar Roland Thuenauer Olga Makshakova Winfried Römer |
| author_facet | Maokai Xu Maria Antonova Pavel Salavei Katharina Illek Ana Valeria Meléndez Ramin Omidvar Roland Thuenauer Olga Makshakova Winfried Römer |
| author_sort | Maokai Xu |
| collection | DOAJ |
| description | Receptor-mediated transcytosis is an elegant and promising strategy for drug delivery across biological barriers. Here, we describe a novel ligand–receptor pair based on a dimeric, engineered derivative of the <i>Pseudomonas aeruginosa</i> lectin LecA, here termed Di-LecA, and the host cell glycosphingolipid Gb3. We characterized the trafficking kinetics and transcytosis efficiencies in polarized Gb3-positive and -negative MDCK cells using mainly immunofluorescence in combination with confocal microscopy. To evaluate the delivery capacity of dimeric LecA chimeras, EGFP was chosen as a fluorescent model protein representing macromolecules, such as antibody fragments, and fused to either the N- or C-terminus of monomeric LecA using recombinant DNA technology. Both LecA/EGFP fusion proteins crossed cellular monolayers in vitro. Of note, the conjugate with EGFP at the N-terminus of LecA (EGFP-LecA) showed a higher release rate than the conjugate with EGFP at the C-terminus (LecA-EGFP). Based on molecular dynamics simulations and cross-linking studies of giant unilamellar vesicles, we speculate that EGFP-LecA tends to be a dimer while LecA-EGFP forms a tetramer. Overall, we confidently propose the dimeric LecA chimeras as transcytotic drug delivery tools through Gb3-positive cellular barriers for future in vivo tests. |
| first_indexed | 2024-03-09T11:27:30Z |
| format | Article |
| id | doaj.art-5be64586fb78427b818bc02dde313482 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T11:27:30Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-5be64586fb78427b818bc02dde3134822023-11-30T23:59:26ZengMDPI AGPharmaceutics1999-49232023-01-0115122510.3390/pharmaceutics15010225Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular BarriersMaokai Xu0Maria Antonova1Pavel Salavei2Katharina Illek3Ana Valeria Meléndez4Ramin Omidvar5Roland Thuenauer6Olga Makshakova7Winfried Römer8Faculty of Biology, University of Freiburg, 79104 Freiburg, GermanyKazan Institute for Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 420111 Kazan, RussiaFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyFaculty of Biology, University of Freiburg, 79104 Freiburg, GermanyReceptor-mediated transcytosis is an elegant and promising strategy for drug delivery across biological barriers. Here, we describe a novel ligand–receptor pair based on a dimeric, engineered derivative of the <i>Pseudomonas aeruginosa</i> lectin LecA, here termed Di-LecA, and the host cell glycosphingolipid Gb3. We characterized the trafficking kinetics and transcytosis efficiencies in polarized Gb3-positive and -negative MDCK cells using mainly immunofluorescence in combination with confocal microscopy. To evaluate the delivery capacity of dimeric LecA chimeras, EGFP was chosen as a fluorescent model protein representing macromolecules, such as antibody fragments, and fused to either the N- or C-terminus of monomeric LecA using recombinant DNA technology. Both LecA/EGFP fusion proteins crossed cellular monolayers in vitro. Of note, the conjugate with EGFP at the N-terminus of LecA (EGFP-LecA) showed a higher release rate than the conjugate with EGFP at the C-terminus (LecA-EGFP). Based on molecular dynamics simulations and cross-linking studies of giant unilamellar vesicles, we speculate that EGFP-LecA tends to be a dimer while LecA-EGFP forms a tetramer. Overall, we confidently propose the dimeric LecA chimeras as transcytotic drug delivery tools through Gb3-positive cellular barriers for future in vivo tests.https://www.mdpi.com/1999-4923/15/1/225Gb3-binding lectinglycosphingolipidtranscytosisdrug carrierprotein engineeringprotein structure |
| spellingShingle | Maokai Xu Maria Antonova Pavel Salavei Katharina Illek Ana Valeria Meléndez Ramin Omidvar Roland Thuenauer Olga Makshakova Winfried Römer Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers Pharmaceutics Gb3-binding lectin glycosphingolipid transcytosis drug carrier protein engineering protein structure |
| title | Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers |
| title_full | Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers |
| title_fullStr | Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers |
| title_full_unstemmed | Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers |
| title_short | Dimeric Lectin Chimeras as Novel Candidates for Gb3-Mediated Transcytotic Drug Delivery through Cellular Barriers |
| title_sort | dimeric lectin chimeras as novel candidates for gb3 mediated transcytotic drug delivery through cellular barriers |
| topic | Gb3-binding lectin glycosphingolipid transcytosis drug carrier protein engineering protein structure |
| url | https://www.mdpi.com/1999-4923/15/1/225 |
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