The Effect of Point Mutations on the Biophysical Properties of an Antimicrobial Peptide: Development of a Screening Protocol for Peptide Stability Screening
| العنوان: | The Effect of Point Mutations on the Biophysical Properties of an Antimicrobial Peptide: Development of a Screening Protocol for Peptide Stability Screening |
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| المؤلفون: | Günther H.J. Peters, Sowmya Indrakumar, Alexander P. Golovanov, Pernille Harris, Inas El Bialy, Werner Streicher, Matja Zalar, Allan Noergaard, Wolfgang Friess, Christin Pohl |
| المصدر: | Molecular Pharmaceutics Pohl, C A, Zalar, M, Bialy, I E, Indrakumar, S, Peters, G H J, Friess, W, Golovanov, A P, Streicher, W W, Noergaard, A & Harris, P 2020, ' The Effect of Point Mutations on the Biophysical Properties of an Antimicrobial Peptide : Development of a Screening Protocol for Peptide Stability Screening ', Molecular Pharmaceutics, vol. 17, no. 9, pp. 3298-3313 . https://doi.org/10.1021/acs.molpharmaceut.0c00406 |
| مصطلحات موضوعية: | protein characterization, Pore Forming Cytotoxic Proteins, Biophysics, Pharmaceutical Science, Peptide, 02 engineering and technology, Protein aggregation, 030226 pharmacology & pharmacy, protein aggregation, Protein–protein interaction, Protein Aggregates, 03 medical and health sciences, pharmaceutical screening, 0302 clinical medicine, Dynamic light scattering, Drug Discovery, medicine, Point Mutation, Thermal stability, chemistry.chemical_classification, Calorimetry, Differential Scanning, Protein Stability, Chemistry, Point mutation, protein engineering, Protein engineering, Hydrogen-Ion Concentration, Plectasin, 021001 nanoscience & nanotechnology, Dynamic Light Scattering, aggregation assessment, peptide screening, Molecular Medicine, protein−protein interactions, Peptides, 0210 nano-technology, medicine.drug |
| الوصف: | Therapeutic peptides and proteins show enormous potential in the pharmaceutical market, but high costs in discovery and development are limiting factors so far. Single or multiple point mutations are commonly introduced in protein drugs to increase their binding affinity or selectivity. They can also induce adverse properties, which might be overlooked in a functional screen, such as a decreased colloidal or thermal stability, leading to problems in later stages of the development. In this study, we address the effect of point mutations on the stability of the 4.4 kDa antimicrobial peptide plectasin, as a case study. We combined a systematic high-throughput biophysical screen of the peptide thermal and colloidal stability using dynamic light scattering and differential scanning calorimetry with structure-based methods including small-angle X-ray scattering, analytical ultracentrifugation, and nuclear magnetic resonance spectroscopy. Additionally, we applied molecular dynamics simulations to link obtained protein stability parameters to the protein's molecular structure. Despite their predicted structural similarities, all four plectasin variants showed substantially different behavior in solution. We observed an increasing propensity of plectasin to aggregate at a higher pH, and the introduced mutations influenced the type of aggregation. Our strategy for systematically assessing the stability and aggregation of protein drugs is generally applicable and is of particular relevance, given the increasing number of protein drugs in development. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1543-8392 1543-8384 |
| DOI: | 10.1021/acs.molpharmaceut.0c00406 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8dacc5f6de055b16d70fda2e85673a8c |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....8dacc5f6de055b16d70fda2e85673a8c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15438392 15438384 |
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| DOI: | 10.1021/acs.molpharmaceut.0c00406 |