A high-throughput integrated microfluidics method enables tyrosine autophosphorylation discovery

Hadas Nevenzal, Meirav Noach-Hirsh, Or Skornik-Bustan, Lev Brio, Efrat Barbiro-Michaely, Yair Glick, Dorit Avrahami, Roxane Lahmi, Amit Tzur, Doron Gerber

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Autophosphorylation of receptor and non-receptor tyrosine kinases is a common molecular switch with broad implications for pathogeneses and therapy of cancer and other human diseases. Technologies for large-scale discovery and analysis of autophosphorylation are limited by the inherent difficulty to distinguish between phosphorylation and autophosphorylation in vivo and by the complexity associated with functional assays of receptors kinases in vitro. Here, we report a method for the direct detection and analysis of tyrosine autophosphorylation using integrated microfluidics and freshly synthesized protein arrays. We demonstrate the efficacy of our platform in detecting autophosphorylation activity of soluble and transmembrane tyrosine kinases, and the dependency of in vitro autophosphorylation assays on membranes. Our method, Integrated Microfluidics for Autophosphorylation Discovery (IMAD), is high-throughput, requires low reaction volumes and can be applied in basic and translational research settings. To our knowledge, it is the first demonstration of posttranslational modification analysis of membrane protein arrays.

Original languageEnglish
Article number42
JournalCommunications Biology
Volume2
Issue number1
DOIs
StatePublished - 1 Dec 2019

Bibliographical note

Publisher Copyright:
© 2019, The Author(s).

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