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Organic & Biomolecular Chemistry
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COMMUNICATION
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solved the problem and the fluorescence intensity generated
under the reaction conditions was confirmed to be directly
proportional to the concentration of peptide 4 (Fig. S7). This
warranted the utility of our system for quantitatively
measuring the permeability even in the presence of a complex
mixture of biological molecules. We conducted the MDCK-II
assay of the same set of peptides (4 and derivatives) using the
FBS-containing medium. Pe values were determined in the
same way with the MDCK-II assay without FBS described above.
As a result, all the peptides exhibited lower permeability
values than on the same assay in a serum-free buffer probably
because the peptides were absorbed on serum proteins (Fig.
3b, red bars). Especially, peptides with hydrophobic residues
such as leucine, valine and phenylalanine at the mutated
residue exhibited much lower values in the presence of FBS.
In summary, we here demonstrated that implementing a
fluorogenic reaction to in vitro permeability assays facilitates
studies of structure-permeability relationship of peptides. The
small size of the terminal alkyne group allowed rapid
measurements of permeability of multiple peptides with little
effects on their permeabilities. Our detection method based
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Conflicts of interest
There are no conflicts to declare.
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Acknowledgements
This work was supported by a Core Research for Evolutional
Science and Technology (CREST) of Molecular Technology (No.
JPMJCR13L4), Japan Science and Technology Agency, grant to
S.S and KAKENHI (No. JP17K13265), Japan Society for the
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