Expression, purification and characterization of human proton-coupled oligopeptide transporter 1 hPEPT1
| العنوان: | Expression, purification and characterization of human proton-coupled oligopeptide transporter 1 hPEPT1 |
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| المؤلفون: | Moazur Rahman, Osman Mirza, Bala K. Prabhala, Magnus Borup Bloch, Thomas Boesen, Heidi A. Ernst, Maria Rafiq, Nadia Mirza, Michael Gajhede, Nanda G. Aduri, Soban Tufail, Nicholas M.I. Taylor |
| المصدر: | Rafiq, M, Ernst, H A, Aduri, N G, Prabhala, B K, Tufail, S, Rahman, M, Bloch, M B, Mirza, N, Taylor, N M I, Boesen, T, Gajhede, M & Mirza, O 2022, ' Expression, purification and characterization of human proton-coupled oligopeptide transporter 1 hPEPT1 ', Protein Expression and Purification, vol. 190, 105990 . https://doi.org/10.1016/j.pep.2021.105990 Rafiq, M, Ernst, H A, Aduri, N G, Prabhala, B K, Tufail, S, Rahman, M, Bloch, M B, Mirza, N, Taylor, N M, Boesen, T, Gajhede, M & Mirza, O 2022, ' Expression, purification and characterization of human proton-coupled oligopeptide transporter 1 hPEPT1 ', Protein Expression and Purification, vol. 190, 105990 . https://doi.org/10.1016/j.pep.2021.105990 |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Peptide transporter, Hot Temperature, PEPTIDE TRANSPORTERS, Gene Expression, Tripeptide, Peptide Transporter 1, Protein purification, Humans, DRUG, Negative-stain electron microscopy, Electrochemical gradient, Thermostability, Oligopeptide, Chemistry, Protein Stability, RECOGNITION, Transporter, Hydrogen-Ion Concentration, Negative stain, Major facilitator superfamily, hPEPT1, Recombinant Proteins, Binding studies, Biochemistry, BETA-LACTAM ANTIBIOTICS, Protein expression, INHIBITORS, FAMILY SLC15, Biotechnology |
| الوصف: | The human peptide transporter hPEPT1 (SLC15A1) is responsible for uptake of dietary di- and tripeptides and a number of drugs from the small intestine by utilizing the proton electrochemical gradient, and hence an important target for peptide-like drug design and drug delivery. hPEPT1 belongs to the ubiquitous major facilitator superfamily that all contain a 12TM core structure, with global conformational changes occurring during the transport cycle. Several bacterial homologues of these transporters have been characterized, providing valuable insight into the transport mechanism of this family. Here we report the overexpression and purification of recombinant hPEPT1 in a detergent-solubilized state. Thermostability profiling of hPEPT1 at different pH values revealed that hPEPT1 is more stable at pH 6 as compared to pH 7 and 8. Micro-scale thermophoresis (MST) confirmed that the purified hPEPT1 was able to bind di- and tripeptides respectively. To assess the in-solution oligomeric state of hPEPT1, negative stain electron microscopy was performed, demonstrating a predominantly monomeric state. |
| وصف الملف: | application/pdf |
| تدمد: | 1096-0279 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::769cbac63b46511a7065e6627c197f41 https://pubmed.ncbi.nlm.nih.gov/34637915 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....769cbac63b46511a7065e6627c197f41 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10960279 |
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