Academic Journal
Investigation of mutations (L41F, F17M, N57E, Y99F_Y134W) effects on the TolAIII-UnaG fluorescence protein's unconjugated bilirubin (UC-BR) binding ability and thermal stability properties
| العنوان: | Investigation of mutations (L41F, F17M, N57E, Y99F_Y134W) effects on the TolAIII-UnaG fluorescence protein's unconjugated bilirubin (UC-BR) binding ability and thermal stability properties |
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| المؤلفون: | Eczacıoğlu, Numan, Ulusu, Yakup, Gökçe, İsa, Lakey, Jeremy H. |
| المساهمون: | Eczacıoğlu, Numan, Ulusu, Yakup |
| بيانات النشر: | Taylor and Francis |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Bilirubin, Fluorescence Protein, UnaG |
| الوصف: | WOS:000678686500001 PubMed ID:34319856 ; The UnaG protein is a ligand (unconjugated bilirubin) dependent fluorescence protein isolated from Unagi freshwater eel larvae and expressed as fusion in heterologous expression systems. Bilirubin is a tetrapyrrole molecule mainly produced from heme catabolism by the destruction of erythrocytes in the body. Bilirubin can cause kernicterus, a serious condition associated with permanent neurological damage in neonates with the passage of brain tissue. Different methods have been developed for plasma bilirubin analysis and quantification. The use of UnaG fluorescence protein triggered by bilirubin has become a new approach in bilirubin studies. In this study, we aimed to investigate the biophysical characterization of ligand interactions with the proteins obtained as a result of mutations (UnaG(Y99F_Y134W), UnaG(N57E), UnaG(L41F), and UnaG(F17M)) on the amino acid sequence of TolAIII-UnaG protein. After the purity levels of the expressed proteins have been analyzed by SDS-PAGE, secondary structures and thermal melting temperatures of the proteins have been examined by circular dichroism spectroscopy. Then determination of excitation and emission points by fluorescence spectroscopy, titration studies have been performed with bilirubin, and dissociation constant was calculated. According to the biophysical characterization studies, UnaG(L41F) has the highest affinity and stability among the mutants. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1082-6068 1532-2297 |
| Relation: | Preparative Biochemistry & Biotechnology; 215Z700; Makale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı; Eczacıoğlu, N., Ulusu, Y., Gökçe, İ., Lakey, J. H. (2021). Investigation of mutations (L41F, F17M, N57E, Y99F_Y134W) effects on the TolAIII-UnaG fluorescence protein's unconjugated bilirubin (UC-BR) binding ability and thermal stability properties, Preparative Biochemistry & Biotechnology, doi:10.1080/10826068.2021.1952597; https://doi.org/10.1080/10826068.2021.1952597; https://hdl.handle.net/11492/5148 |
| DOI: | 10.1080/10826068.2021.1952597 |
| الاتاحة: | https://hdl.handle.net/11492/5148 https://doi.org/10.1080/10826068.2021.1952597 |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.69AB5849 |
| قاعدة البيانات: | BASE |
| تدمد: | 10826068 15322297 |
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| DOI: | 10.1080/10826068.2021.1952597 |