Split Intein-Mediated Protein Ligation for detecting protein-protein interactions and their inhibition

Zhong Yao, Farzaneh Aboualizadeh, Jason Kroll, Indira Akula, Jamie Snider, Anna Lyakisheva, Priscilla Tang, Max Kotlyar, Igor Jurisica, Mike Boxem and Igor Stagljar ()
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Zhong Yao: University of Toronto
Farzaneh Aboualizadeh: University of Toronto
Jason Kroll: Utrecht University
Indira Akula: University of Toronto
Jamie Snider: University of Toronto
Anna Lyakisheva: University of Toronto
Priscilla Tang: University of Toronto
Max Kotlyar: University Health Network
Igor Jurisica: University Health Network
Mike Boxem: Utrecht University
Igor Stagljar: University of Toronto

Nature Communications, 2020, vol. 11, issue 1, 1-14

Abstract: Abstract Here, to overcome many limitations accompanying current available methods to detect protein-protein interactions (PPIs), we develop a live cell method called Split Intein-Mediated Protein Ligation (SIMPL). In this approach, bait and prey proteins are respectively fused to an intein N-terminal fragment (IN) and C-terminal fragment (IC) derived from a re-engineered split intein GP41-1. The bait/prey binding reconstitutes the intein, which splices the bait and prey peptides into a single intact protein that can be detected by regular protein detection methods such as Western blot analysis and ELISA, serving as readouts of PPIs. The method is robust and can be applied not only in mammalian cell lines but in animal models such as C. elegans. SIMPL demonstrates high sensitivity and specificity, and enables exploration of PPIs in different cellular compartments and tracking of kinetic interactions. Additionally, we establish a SIMPL ELISA platform that enables high-throughput screening of PPIs and their inhibitors.

Date: 2020
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DOI: 10.1038/s41467-020-16299-1

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