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Unstable Protein Purification Through the Formation of Stable Complexes.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1764, pp. 315-328. - Publication Year :
- 2018
-
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.
- Subjects :
- Adaptor Proteins, Signal Transducing chemistry
Adaptor Proteins, Signal Transducing metabolism
HIV Integrase chemistry
HIV Integrase metabolism
Humans
Nuclear Receptor Coactivator 2 chemistry
Nuclear Receptor Coactivator 2 metabolism
Protein Binding
Receptors, Cytoplasmic and Nuclear chemistry
Receptors, Cytoplasmic and Nuclear metabolism
Receptors, Glucocorticoid chemistry
Receptors, Glucocorticoid metabolism
Transcription Factors chemistry
Transcription Factors metabolism
Adaptor Proteins, Signal Transducing isolation & purification
Chromatography, Affinity methods
HIV Integrase isolation & purification
Nuclear Receptor Coactivator 2 isolation & purification
Protein Interaction Domains and Motifs
Receptors, Cytoplasmic and Nuclear isolation & purification
Receptors, Glucocorticoid isolation & purification
Transcription Factors isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1764
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 29605924
- Full Text :
- https://doi.org/10.1007/978-1-4939-7759-8_20