@inbook{421626ca669649e7bc03b30f3703e5ed,
title = "An Interactome Assay for Detecting Interactions between Extracellular Domains of Receptor Kinases",
abstract = "Interactions between extracellular domains (ECDs) are crucial for many physiological processes in the cell, most importantly perception of its environment. However, studying these often-transient interactions can be challenging. Here we describe a method that allows for in vitro detection of extracellular domain interactions through an oligomerization-based cell surface interaction (CSI) assay. In a CSI, bait- and prey-tagged proteins are produced and secreted by Drosophila S2 cells to ensure proper folding and post-translational modifications. Subsequently, Bait (FC fragment) and Prey (pentamer domain and alkaline phosphatase) tags allow the detection of interactions in protein A-coated 96 wells plates through a colorimetric readout. Due to the easy detection of interactions this approach can be used for high-throughput screening and mapping of extracellular interaction networks.",
keywords = "ECDs, High-throughput screen, Interactome assay, Protein–protein interactions, Receptor kinases, S2 cell protein expression, Signaling",
author = "Jente Stouthamer and Sergio Martin-Ramirez and Elwira Smakowska-Luzan",
year = "2023",
month = jul,
day = "15",
doi = "10.1007/978-1-0716-3327-4_18",
language = "English",
isbn = "9781071633298",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "193--204",
editor = "S. Mukhtar",
booktitle = "Methods in Molecular Biology",
address = "United States",
}