Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells.
Aggregation inhibitor
Luciferase
Luminescence
Protein solubility
Split-protein reassembly
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2019
2019
Historique:
pmc-release:
09
03
2020
entrez:
4
6
2019
pubmed:
4
6
2019
medline:
23
2
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31155065
pii: S0076-6879(19)30022-9
doi: 10.1016/bs.mie.2019.02.003
pmc: PMC6567984
mid: NIHMS1033950
pii:
doi:
Substances chimiques
Luminescent Agents
0
Protein Aggregates
0
Proteins
0
Luciferases
EC 1.13.12.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
55-66Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM119751
Pays : United States
Informations de copyright
© 2019 Elsevier Inc. All rights reserved.
Références
ChemMedChem. 2007 Dec;2(12):1674-92
pubmed: 17952881
Protein Sci. 1999 Sep;8(9):1908-11
pubmed: 10493593
Mol Biosyst. 2016 Oct 20;12(10):2984-7
pubmed: 27483434
Protein Sci. 1993 Mar;2(3):404-10
pubmed: 8453378
ACS Chem Biol. 2016 Jan 15;11(1):132-8
pubmed: 26492083
Methods Enzymol. 1999;309:3-25
pubmed: 10507013
Biochem Biophys Res Commun. 2016 Jan 29;470(1):88-93
pubmed: 26746005
Annu Rev Biochem. 2017 Jun 20;86:27-68
pubmed: 28498720
Annu Rev Biochem. 2006;75:333-66
pubmed: 16756495
Nat Biotechnol. 2001 Feb;19(2):131-6
pubmed: 11175726
ACS Chem Biol. 2012 Nov 16;7(11):1848-57
pubmed: 22894855
ACS Chem Biol. 2006 Aug 22;1(7):461-9
pubmed: 17168524
Nat Biotechnol. 1999 Jul;17(7):691-5
pubmed: 10404163