Quality Control of Proteins Solubilized from Inclusion Bodies.
Circular dichroism spectroscopy
Dynamic light scattering
Inclusion body
Insoluble protein
Multilamellar vesicles
Recombinant protein
Small unilamellar vesicles
Thermal stability
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
28
1
2022
pubmed:
29
1
2022
medline:
31
3
2022
Statut:
ppublish
Résumé
Despite substantial development of production and purification protocols for heterologous recombinant proteins, some proteins are difficult to produce or, when produced, are accumulated in inclusion bodies (IBs). Nondenaturing protocols can be used to recover the entrapped protein from these protein aggregates. In this chapter, we provide a detailed procedure to analyze the physicochemical properties of one of those proteins produced in prokaryotic expression systems. Serum amyloid A3 (SAA3) was recovered from inclusion bodies (IBs) and its secondary structure associated to thermal stability and size was determined by circular dichroism (CD) and dynamic light scattering (DLS), respectively. These techniques were also applied to evaluate the SAA3 interaction with model membranes. These results show the importance of the structural analysis of proteins released from inclusion bodies under nondenaturing procedures, although similar approaches can be extended to any type of recombinant protein preparation.
Identifiants
pubmed: 35089575
doi: 10.1007/978-1-0716-1859-2_28
doi:
Substances chimiques
Recombinant Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
469-477Informations de copyright
© 2022. Springer Science+Business Media, LLC, part of Springer Nature.
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