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@ARTICLE{Xu:614232,
author = {Xu, Maokai and Antonova, Maria and Salavei, Pavel and
Illek, Katharina and Meléndez, Ana Valeria and Omidvar,
Ramin and Thuenauer, Roland and Makshakova, Olga and Roemer,
Winfried},
title = {{D}imeric {L}ectin {C}himeras as {N}ovel {C}andidates for
{G}b3-{M}ediated {T}ranscytotic {D}rug {D}elivery through
{C}ellular {B}arriers},
journal = {Pharmaceutics},
volume = {15},
number = {1},
issn = {1999-4923},
address = {Basel},
publisher = {MDPI},
reportid = {PUBDB-2024-05772},
pages = {225},
year = {2023},
abstract = {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 Pseudomonas aeruginosa
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.Keywords:Gb3-binding lectin;
glycosphingolipid; transcytosis; drug carrier; protein
engineering; protein structure; valency; accelerated
molecular dynamics},
cin = {CSSB-CF-ALFM},
ddc = {610},
cid = {I:(DE-H253)CSSB-CF-ALFM-20210629},
pnm = {899 - ohne Topic (POF4-899) / DFG project
G:(GEPRIS)39236281 - EXC 294: BIOSS Zentrum für Biologische
Signalstudien - von der Analyse zur Synthese (39236281) /
DFG project G:(GEPRIS)390939984 - EXC 2189: CIBSS - Centre
for Integrative Biological Signalling Studies (390939984)},
pid = {G:(DE-HGF)POF4-899 / G:(GEPRIS)39236281 /
G:(GEPRIS)390939984},
experiment = {EXP:(DE-MLZ)NOSPEC-20140101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:36678854},
UT = {WOS:000918781700001},
doi = {10.3390/pharmaceutics15010225},
url = {https://bib-pubdb1.desy.de/record/614232},
}