Unstable Protein Purification Through the Formation of Stable Complexes
| Autor: | Oyindamola Oladosu, Marc Ruff, Sylvia Eiler, Benoit Maillot, Karine Pradeau Aubreton, Julien Batisse, Nicolas Lévy |
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| Přispěvatelé: | Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Protein family Chemistry medicine.medical_treatment Combinatorial chemistry Pre-integration complex Steroid 03 medical and health sciences 030104 developmental biology Protein purification Protein biosynthesis medicine Protein complex formation [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Functional studies Solubility ComputingMilieux_MISCELLANEOUS |
| Zdroj: | Methods in Molecular Biology Methods in Molecular Biology, pp.315-328, 2018, ⟨10.1007/978-1-4939-7759-8_20⟩ Protein Complex Assembly ISBN: 9781493977581 |
| DOI: | 10.1007/978-1-4939-7759-8_20⟩ |
| Popis: | Purification of proteins containing disordered regions and participating in transient complexes is often challenging because of the small amounts available after purification, their heterogeneity, instability, and/or poor solubility. To circumvent these difficulties, we set up a methodology that enables the production of stable complexes in large amounts for structural and functional studies. In this chapter, we describe the methodology used to establish the best cell culture conditions and buffer compositions to optimize soluble protein production and their stabilization through protein complex formation. Two examples of challenging protein families are described, namely, the human steroid nuclear receptors and the HIV-1 pre-integration complexes. |
| Databáze: | OpenAIRE |
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