دورية أكاديمية
Bacterial expression and purification of vertebrate odorant-binding proteins.
| العنوان: | Bacterial expression and purification of vertebrate odorant-binding proteins. |
|---|---|
| المؤلفون: | Brulé M; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France., Glaz M; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France., Belloir C; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France., Poirier N; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France., Moitrier L; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France., Neiers F; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France., Briand L; Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France. Electronic address: loic.briand@inrae.fr. |
| المصدر: | Methods in enzymology [Methods Enzymol] 2020; Vol. 642, pp. 125-150. Date of Electronic Publication: 2020 May 30. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 0212271 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1557-7988 (Electronic) Linking ISSN: 00766879 NLM ISO Abbreviation: Methods Enzymol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: New York, Academic Press. |
| مواضيع طبية MeSH: | Receptors, Odorant*/genetics , Receptors, Odorant*/metabolism, Animals ; Carrier Proteins ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Insect Proteins ; Odorants ; Swine ; Vertebrates/metabolism |
| مستخلص: | Vertebrate odorant-binding proteins (OBPs) are small soluble proteins abundantly secreted in the olfactory mucus of many animal species, including humans. Vertebrate OBPs reversibly bind odorant molecules with micromolar range affinities. Although their physiological role is not clearly understood, OBPs are proposed to carry airborne odorants toward membrane olfactory receptors through the nasal mucus. Measurements of odorant-OBP interactions and structural studies require a large amount of pure OBPs devoid of ligands. The bacterial expression system is the first choice for expressing vertebrate OBPs used in our laboratory and others. This system generally produces OBPs in large amounts without major problems. In this chapter, we describe the milligram-scale production of recombinant pig OBP1 (pOBP1) in E. coli. The different steps of expression and purification are presented and discussed. Protocols for secondary structures investigation by circular dichroism and binding properties of the recombinant protein are also provided. More generally, these approaches can be used to produce and characterize any vertebrate OBPs for use in functional and structural studies. (© 2020 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Affinity; Circular dichroism; Fluorescent assay; Isothermal microcalorimetry; Lipocalin; Odorant; Odorant-binding protein; Purification |
| المشرفين على المادة: | 0 (Carrier Proteins) 0 (Insect Proteins) 0 (Receptors, Odorant) |
| تواريخ الأحداث: | Date Created: 20200824 Date Completed: 20210623 Latest Revision: 20210623 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/bs.mie.2020.05.002 |
| PMID: | 32828250 |
| قاعدة البيانات: | MEDLINE |
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