Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells.
Methods Enzymol
; 622: 55-66, 2019.
Article
en En
| MEDLINE
| ID: mdl-31155065
ABSTRACT
Protein misfolding and aggregation is now recognized as a hallmark of numerous human diseases. Standard bioanalytical techniques for monitoring protein aggregation generally rely on small molecules that provide an optical readout of fibril formation. While these methods have been useful for mechanistic studies, additional approaches are required to probe the equilibrium between soluble and insoluble protein within living systems. Such approaches could provide platforms for the identification of inhibitors of protein aggregation as well as a means to investigate the effect of mutations on protein aggregation in model systems. In this chapter, we provide detailed protocols for employing split-NanoLuc luciferase (Nluc) fragments to monitor changes in protein solubility in bacterial and mammalian cells. This sensitive luminesce-based assay can report upon changes in protein solubility induced by inhibitors and disease-relevant mutations.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Proteínas
/
Sustancias Luminiscentes
/
Agregado de Proteínas
/
Luciferasas
/
Mediciones Luminiscentes
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Methods Enzymol
Año:
2019
Tipo del documento:
Article
País de afiliación:
Estados Unidos