Self-Assembling NanoLuc Luciferase Fragments as Probes for Protein Aggregation in Living Cells

Jia Zhao, Travis J. Nelson, Quyen Vu, Tiffany Truong, Cliff I. Stains

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Given the clear role of protein aggregation in human disease, there is a critical need for assays capable of quantifying protein aggregation in living systems. We hypothesized that the inherently low background and biocompatibility of luminescence signal readouts could provide a potential solution to this problem. Herein, we describe a set of self-assembling NanoLuc luciferase (Nluc) fragments that produce a tunable luminescence readout that is dependent upon the solubility of a target protein fused to the N-terminal Nluc fragment. To demonstrate this approach, we employed this assay in bacteria to assess mutations known to disrupt amyloid-beta (Aβ) aggregation as well as disease-relevant mutations associated with familial Alzheimers diseases. The luminescence signal from these experiments correlates with the reported aggregation potential of these Aβ mutants and reinforces the increased aggregation potential of disease-relevant mutations in Aβ1-42. To further demonstrate the utility of this approach, we show that the effect of small molecule inhibitors on Aβ aggregation can be monitored using this system. In addition, we demonstrate that aggregation assays can be ported into mammalian cells. Taken together, these results indicate that this platform could be used to rapidly screen for mutations that influence protein aggregation as well as inhibitors of protein aggregation. This method offers a novel, genetically encodable luminescence readout of protein aggregation in living cells.

Original languageEnglish (US)
Pages (from-to)132-138
Number of pages7
JournalACS chemical biology
Volume11
Issue number1
DOIs
StatePublished - Jan 15 2016

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

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