Analysis of Protein-Protein Interactions by Split Luciferase Complementation Assay

Yuekun Lang; Zhong Li; Hongmin Li

doi:10.1002/cptx.90

Analysis of Protein-Protein Interactions by Split Luciferase Complementation Assay

Yuekun Lang, Zhong Li, Hongmin Li

Research output: Contribution to journal › Article › peer-review

11 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 language	English (US)
Article number	e90
Journal	Current Protocols in Toxicology
Volume	82
Issue number	1
DOIs	https://doi.org/10.1002/cptx.90
State	Published - Dec 1 2019
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

Toxicology

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10.1002/cptx.90

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title = "Analysis of Protein-Protein Interactions by Split Luciferase Complementation Assay",

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.",

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AB - 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.

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Analysis of Protein-Protein Interactions by Split Luciferase Complementation Assay

Abstract

Keywords

ASJC Scopus subject areas

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