Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells

Travis J. Nelson, Jia Zhao, Cliff I. Stains

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsArun K. Shukla
PublisherAcademic Press Inc.
Pages55-66
Number of pages12
ISBN (Print)9780128181195
DOIs
StatePublished - 2019

Publication series

NameMethods in Enzymology
Volume622
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Luminescent Proteins
Luciferases
Solubility
Cells
Agglomeration
Proteins
Mutation
Assays
Molecules
Monitoring

Keywords

  • Aggregation inhibitor
  • Luciferase
  • Luminescence
  • Protein solubility
  • Split-protein reassembly

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Nelson, T. J., Zhao, J., & Stains, C. I. (2019). Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells. In A. K. Shukla (Ed.), Methods in Enzymology (pp. 55-66). (Methods in Enzymology; Vol. 622). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2019.02.003

Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells. / Nelson, Travis J.; Zhao, Jia; Stains, Cliff I.

Methods in Enzymology. ed. / Arun K. Shukla. Academic Press Inc., 2019. p. 55-66 (Methods in Enzymology; Vol. 622).

Research output: Chapter in Book/Report/Conference proceedingChapter

Nelson, TJ, Zhao, J & Stains, CI 2019, Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells. in AK Shukla (ed.), Methods in Enzymology. Methods in Enzymology, vol. 622, Academic Press Inc., pp. 55-66. https://doi.org/10.1016/bs.mie.2019.02.003
Nelson TJ, Zhao J, Stains CI. Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells. In Shukla AK, editor, Methods in Enzymology. Academic Press Inc. 2019. p. 55-66. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2019.02.003
Nelson, Travis J. ; Zhao, Jia ; Stains, Cliff I. / Utilizing split-NanoLuc luciferase fragments as luminescent probes for protein solubility in living cells. Methods in Enzymology. editor / Arun K. Shukla. Academic Press Inc., 2019. pp. 55-66 (Methods in Enzymology).
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