Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion
Rapid digestion by proteases limits the application of peptides as therapeutics. One strategy to increase the proteolytic stability of peptides is the modification with fluorinated amino acids. This study presents a systematic investigation of the effects of fluorinated leucine and isoleucine deriva...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Beilstein-Institut
2017-12-01
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| Series: | Beilstein Journal of Organic Chemistry |
| Subjects: | |
| Online Access: | https://doi.org/10.3762/bjoc.13.279 |
| _version_ | 1818956369758257152 |
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| author | Susanne Huhmann Anne-Katrin Stegemann Kristin Folmert Damian Klemczak Johann Moschner Michelle Kube Beate Koksch |
| author_facet | Susanne Huhmann Anne-Katrin Stegemann Kristin Folmert Damian Klemczak Johann Moschner Michelle Kube Beate Koksch |
| author_sort | Susanne Huhmann |
| collection | DOAJ |
| description | Rapid digestion by proteases limits the application of peptides as therapeutics. One strategy to increase the proteolytic stability of peptides is the modification with fluorinated amino acids. This study presents a systematic investigation of the effects of fluorinated leucine and isoleucine derivatives on the proteolytic stability of a peptide that was designed to comprise substrate specificities of different proteases. Therefore, leucine, isoleucine, and their side-chain fluorinated variants were site-specifically incorporated at different positions of this peptide resulting in a library of 13 distinct peptides. The stability of these peptides towards proteolysis by α-chymotrypsin, pepsin, proteinase K, and elastase was studied, and this process was followed by an FL-RP-HPLC assay in combination with mass spectrometry. In a few cases, we observed an exceptional increase in proteolytic stability upon introduction of the fluorine substituents. The opposite phenomenon was observed in other cases, and this may be explained by specific interactions of fluorinated residues with the respective enzyme binding sites. Noteworthy is that 5,5,5-trifluoroisoleucine is able to significantly protect peptides from proteolysis by all enzymes included in this study when positioned N-terminal to the cleavage site. These results provide valuable information for the application of fluorinated amino acids in the design of proteolytically stable peptide-based pharmaceuticals. |
| first_indexed | 2024-12-20T10:52:52Z |
| format | Article |
| id | doaj.art-33c87508740d4c4d9a88456046cc4f3e |
| institution | Directory Open Access Journal |
| issn | 1860-5397 |
| language | English |
| last_indexed | 2024-12-20T10:52:52Z |
| publishDate | 2017-12-01 |
| publisher | Beilstein-Institut |
| record_format | Article |
| series | Beilstein Journal of Organic Chemistry |
| spelling | doaj.art-33c87508740d4c4d9a88456046cc4f3e2022-12-21T19:43:13ZengBeilstein-InstitutBeilstein Journal of Organic Chemistry1860-53972017-12-011312869288210.3762/bjoc.13.2791860-5397-13-279Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestionSusanne Huhmann0Anne-Katrin Stegemann1Kristin Folmert2Damian Klemczak3Johann Moschner4Michelle Kube5Beate Koksch6Department of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyDepartment of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyDepartment of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyDepartment of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyDepartment of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyDepartment of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyDepartment of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, GermanyRapid digestion by proteases limits the application of peptides as therapeutics. One strategy to increase the proteolytic stability of peptides is the modification with fluorinated amino acids. This study presents a systematic investigation of the effects of fluorinated leucine and isoleucine derivatives on the proteolytic stability of a peptide that was designed to comprise substrate specificities of different proteases. Therefore, leucine, isoleucine, and their side-chain fluorinated variants were site-specifically incorporated at different positions of this peptide resulting in a library of 13 distinct peptides. The stability of these peptides towards proteolysis by α-chymotrypsin, pepsin, proteinase K, and elastase was studied, and this process was followed by an FL-RP-HPLC assay in combination with mass spectrometry. In a few cases, we observed an exceptional increase in proteolytic stability upon introduction of the fluorine substituents. The opposite phenomenon was observed in other cases, and this may be explained by specific interactions of fluorinated residues with the respective enzyme binding sites. Noteworthy is that 5,5,5-trifluoroisoleucine is able to significantly protect peptides from proteolysis by all enzymes included in this study when positioned N-terminal to the cleavage site. These results provide valuable information for the application of fluorinated amino acids in the design of proteolytically stable peptide-based pharmaceuticals.https://doi.org/10.3762/bjoc.13.279fluorinated amino acidshexafluoroleucinepeptide drugsprotease stabilitytrifluoroisoleucine |
| spellingShingle | Susanne Huhmann Anne-Katrin Stegemann Kristin Folmert Damian Klemczak Johann Moschner Michelle Kube Beate Koksch Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion Beilstein Journal of Organic Chemistry fluorinated amino acids hexafluoroleucine peptide drugs protease stability trifluoroisoleucine |
| title | Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion |
| title_full | Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion |
| title_fullStr | Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion |
| title_full_unstemmed | Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion |
| title_short | Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion |
| title_sort | position dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion |
| topic | fluorinated amino acids hexafluoroleucine peptide drugs protease stability trifluoroisoleucine |
| url | https://doi.org/10.3762/bjoc.13.279 |
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