Analysis of Protein-Protein Interactions by Split Luciferase Complementation Assay

Yuekun Lang, Zhong Li, Hongmin Li

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Protein-protein interactions are important in human disease. Developing and refining tools to understand physical contacts between signaling proteins is crucial. This article describes a split luciferase complementation (SLC) method designed to discover inhibitors of protein-protein interaction. Different fusion proteins with split luciferase are constructed, expressed, and purified, and then assessed to determine the best pair that generates the strongest luminescence. SLC specificity and affinity are further confirmed. Step-by-step instructions are provided for performing these assays using the NS2B-NS3 interaction as an example. NS2B is an essential cofactor for flaviviral NS3 protease function. Advantages and disadvantages of these assays are further discussed.

Original languageEnglish (US)
Article numbere90
JournalCurrent Protocols in Toxicology
Volume82
Issue number1
DOIs
StatePublished - Dec 1 2019
Externally publishedYes

Keywords

  • NS2B-NS3
  • flaviviral protease
  • protein-protein interaction
  • split luciferase complementation

ASJC Scopus subject areas

  • Toxicology

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