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

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.

journal_name

Nat Commun

journal_title

Nature communications

authors

Yao Z,Aboualizadeh F,Kroll J,Akula I,Snider J,Lyakisheva A,Tang P,Kotlyar M,Jurisica I,Boxem M,Stagljar I

doi

10.1038/s41467-020-16299-1

subject

Has Abstract

pub_date

2020-05-15 00:00:00

pages

2440

issue

1

issn

2041-1723

pii

10.1038/s41467-020-16299-1

journal_volume

11

pub_type

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