Chapter title |
Unstable Protein Purification Through the Formation of Stable Complexes
|
---|---|
Chapter number | 20 |
Book title |
Protein Complex Assembly
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7759-8_20 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7758-1, 978-1-4939-7759-8
|
Authors |
Sylvia Eiler, Nicolas Levy, Benoit Maillot, Julien Batisse, Karine Pradeau Aubreton, Oyindamola Oladosu, Marc Ruff |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 11 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 4 | 36% |
Student > Bachelor | 1 | 9% |
Student > Ph. D. Student | 1 | 9% |
Unknown | 5 | 45% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 2 | 18% |
Agricultural and Biological Sciences | 2 | 18% |
Chemistry | 1 | 9% |
Medicine and Dentistry | 1 | 9% |
Unknown | 5 | 45% |