| Literature DB >> 31250310 |
Rina Barouch-Bentov1, Yves Jacob2, Shirit Einav3,4.
Abstract
The endosomal sorting complex required for transport (ESCRT) machinery comprises five complexes that act sequentially to recruit and cluster transmembrane cargo proteins (ESCRT-0), drive membrane curving (ESCRT-I and II), catalyze fission of cargo-containing vesicles (ESCRT-III and VPS/VTA1), and disassemble and recycle the ESCRT-III complex (VPS/VTA1). Since interactions between ESCRT components and cellular or microbial proteins are typically weak, transient, and involve membrane proteins, they are often difficult to study by standard technologies. Here, we describe the utility of high-throughput protein-fragment complementation assays based on the reconstitution of a split luciferase reporter to screen for interactions between any protein and a library of ESCRT proteins in mammalian cells and provide a detailed protocol for these assays.Entities:
Keywords: ESCRT; Protein-fragment complementation assays; Protein-protein interactions; Proteomics
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Year: 2019 PMID： 31250310 DOI： 10.1007/978-1-4939-9492-2_21
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745