Abstract
Here we describe the biochemical assay for ATE1-mediated arginylation in microplate format, which can be applied to high-throughput screens for identification of small-molecule inhibitors and activators of ATE1, high-volume analysis of ATE1 substrates, and other similar applications. Originally, we have applied this screen to a library of 3280 compounds and identified two compounds which specifically affect ATE1-regulated processes in vitro and in vivo. The assay is based on in vitro ATE1-mediated arginylation of beta-actin’s N-terminal peptide, but it can also be applied using other ATE1 substrates.
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References
Saha S, Wang J, Buckley B, Wang Q, Lilly B, Chernov M, Kashina A (2012) Small molecule inhibitors of arginyltransferase regulate arginylation-dependent protein degradation, cell motility, and angiogenesis. Biochem Pharmacol 83(7):866–873
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© 2015 Springer Science+Business Media New York
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Saha, S., Wang, J., Kashina, A.S. (2015). High-Throughput Arginylation Assay in Microplate Format. In: Kashina, A. (eds) Protein Arginylation. Methods in Molecular Biology, vol 1337. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2935-1_11
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DOI: https://doi.org/10.1007/978-1-4939-2935-1_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2934-4
Online ISBN: 978-1-4939-2935-1
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