Small-molecule screening assay for mono-adp-ribosyltransferases
Haikarainen, Teemu; Murthy, Sudarshan; Maksimainen, Mirko M.; Lehtiö, Lari (2018-08-11)
Haikarainen T., Murthy S., Maksimainen M.M., Lehtiö L. (2018) Small-Molecule Screening Assay for Mono-ADP-Ribosyltransferases. In: Chang P. (eds) ADP-ribosylation and NAD+ Utilizing Enzymes. Methods in Molecular Biology, vol 1813. Humana Press, New York, NY
© Springer Science+Business Media, LLC, part of Springer Nature 2018. This is a post-peer-review, pre-copyedit version of an article published in ADP-ribosylation and NAD+ Utilizing Enzymes. The final authenticated version is available online at: https://doi.org/10.1007/978-1-4939-8588-3_16.
https://rightsstatements.org/vocab/InC/1.0/
https://urn.fi/URN:NBN:fi-fe201902226030
Tiivistelmä
Abstract
Mono-ADP-ribosyltransferases of the PARP/ARTD enzyme family are enzymes catalyzing the transfer of a single ADP-ribose unit to target proteins. The enzymes have various roles in vital cellular processes such as DNA repair and transcription, and many of the enzymes are linked to cancer-relevant functions. Thus inhibition of the enzymes is a potential way to discover and develop new drugs against cancer. Here we describe an activity-based screening assay for mono-ADP-ribosyltransferases. The assay utilizes the natural substrate of the enzymes, NAD+, and it is based on chemically converting the leftover substrate to a fluorophore and measuring its relative concentration after the enzymatic reaction. The assay is homogenous, robust, and cost-effective and, most importantly, applicable to mono-ADP-ribosyltransferases as well as poly-ADP-ribosyltransferases for screening of small-molecule inhibitors against the enzymes.
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