Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25
Peptide oligomerization dynamics affects peptide structure, activity, and pharmacodynamic properties. The thrombin C-terminal peptide, TCP-25 (GKYGFYTHVFRLKKWIQKVIDQFGE), is currently in preclinical development for improved wound healing and infection prevention. It exhibits turbidity when formulate...
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| Format: | Article |
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MDPI AG
2020-11-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/11/1572 |
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| author | Ganna Petruk Jitka Petrlova Firdaus Samsudin Rita Del Giudice Peter J. Bond Artur Schmidtchen |
| author_facet | Ganna Petruk Jitka Petrlova Firdaus Samsudin Rita Del Giudice Peter J. Bond Artur Schmidtchen |
| author_sort | Ganna Petruk |
| collection | DOAJ |
| description | Peptide oligomerization dynamics affects peptide structure, activity, and pharmacodynamic properties. The thrombin C-terminal peptide, TCP-25 (GKYGFYTHVFRLKKWIQKVIDQFGE), is currently in preclinical development for improved wound healing and infection prevention. It exhibits turbidity when formulated at pH 7.4, particularly at concentrations of 0.3 mM or more. We used biochemical and biophysical approaches to explore whether the peptide self-associates and forms oligomers. The peptide showed a dose-dependent increase in turbidity as well as α-helical structure at pH 7.4, a phenomenon not observed at pH 5.0. By analyzing the intrinsic tryptophan fluorescence, we demonstrate that TCP-25 is more stable at high concentrations (0.3 mM) when exposed to high temperatures or a high concentration of denaturant agents, which is compatible with oligomer formation. The denaturation process was reversible above 100 µM of peptide. Dynamic light scattering demonstrated that TCP-25 oligomerization is sensitive to changes in pH, time, and temperature. Computational modeling with an active 18-mer region of TCP-25 showed that the peptide can form pH-dependent higher-order end-to-end oligomers and micelle-like structures, which is in agreement with the experimental data. Thus, TCP-25 exhibits pH- and temperature-dependent dynamic changes involving helical induction and reversible oligomerization, which explains the observed turbidity of the pharmacologically developed formulation. |
| first_indexed | 2024-03-10T14:44:18Z |
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| id | doaj.art-d5d1c3c78bc64686a51b40fc3d7fd894 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T14:44:18Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-d5d1c3c78bc64686a51b40fc3d7fd8942023-11-20T21:31:41ZengMDPI AGBiomolecules2218-273X2020-11-011011157210.3390/biom10111572Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25Ganna Petruk0Jitka Petrlova1Firdaus Samsudin2Rita Del Giudice3Peter J. Bond4Artur Schmidtchen5Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University, 22241 Lund, SwedenDivision of Dermatology and Venereology, Department of Clinical Sciences, Lund University, 22241 Lund, SwedenBioinformatics Institute (BII), Agency for Science, Technology and Research (A*STAR), Singapore 138671, SingaporeResearch Centre for Biointerfaces, Department of Biomedical Sciences and Biofilms, Malmö University, 20506 Malmö, SwedenBioinformatics Institute (BII), Agency for Science, Technology and Research (A*STAR), Singapore 138671, SingaporeDivision of Dermatology and Venereology, Department of Clinical Sciences, Lund University, 22241 Lund, SwedenPeptide oligomerization dynamics affects peptide structure, activity, and pharmacodynamic properties. The thrombin C-terminal peptide, TCP-25 (GKYGFYTHVFRLKKWIQKVIDQFGE), is currently in preclinical development for improved wound healing and infection prevention. It exhibits turbidity when formulated at pH 7.4, particularly at concentrations of 0.3 mM or more. We used biochemical and biophysical approaches to explore whether the peptide self-associates and forms oligomers. The peptide showed a dose-dependent increase in turbidity as well as α-helical structure at pH 7.4, a phenomenon not observed at pH 5.0. By analyzing the intrinsic tryptophan fluorescence, we demonstrate that TCP-25 is more stable at high concentrations (0.3 mM) when exposed to high temperatures or a high concentration of denaturant agents, which is compatible with oligomer formation. The denaturation process was reversible above 100 µM of peptide. Dynamic light scattering demonstrated that TCP-25 oligomerization is sensitive to changes in pH, time, and temperature. Computational modeling with an active 18-mer region of TCP-25 showed that the peptide can form pH-dependent higher-order end-to-end oligomers and micelle-like structures, which is in agreement with the experimental data. Thus, TCP-25 exhibits pH- and temperature-dependent dynamic changes involving helical induction and reversible oligomerization, which explains the observed turbidity of the pharmacologically developed formulation.https://www.mdpi.com/2218-273X/10/11/1572oligomerizationthrombinpeptide self-assemblyantimicrobial peptidepH- and/or concentration-sensitive oligomerizationTCP-25 |
| spellingShingle | Ganna Petruk Jitka Petrlova Firdaus Samsudin Rita Del Giudice Peter J. Bond Artur Schmidtchen Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25 Biomolecules oligomerization thrombin peptide self-assembly antimicrobial peptide pH- and/or concentration-sensitive oligomerization TCP-25 |
| title | Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25 |
| title_full | Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25 |
| title_fullStr | Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25 |
| title_full_unstemmed | Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25 |
| title_short | Concentration- and pH-Dependent Oligomerization of the Thrombin-Derived C-Terminal Peptide TCP-25 |
| title_sort | concentration and ph dependent oligomerization of the thrombin derived c terminal peptide tcp 25 |
| topic | oligomerization thrombin peptide self-assembly antimicrobial peptide pH- and/or concentration-sensitive oligomerization TCP-25 |
| url | https://www.mdpi.com/2218-273X/10/11/1572 |
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