@inbook{80eb64472dc648a2ab6a4c8eab123b29,
title = "A pyrophosphatase-coupled assay to monitor the NTase activity of cGAS",
abstract = "Cyclic GMP-AMP synthase, cGAS, converts ATP and GTP into a cyclic dinucleotide second messenger, cyclic GMP-AMP or cGAMP, through its enzymatic, nucleotidyl transferase (NTase) activity. Although many methods are available to directly measure cGAMP production, these assays often have high cost of implementation and/or experimental limitations. This chapter details how to implement an alternative approach that is relatively inexpensive, accurate and medium-throughput. The assay measures cGAS NTase activity by quantifying pyrophosphate production, a byproduct of the cGAS reaction. A coupling enzyme, pyrophosphatase, catalyzes the hydrolysis of pyrophosphate into inorganic phosphate, which enables facile detection of cGAS activity through conventional phosphomolybdate-malachite green absorbance methodology. This method is amenable for conventional steady-state kinetic measurements as well as high-throughput compound screening.",
keywords = "Nucleotidyl transferase (NTase), PPase, Pyrophosphatase, Steady-state, cGAS",
author = "Richard Hooy and Jungsan Sohn",
note = "Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
doi = "10.1016/bs.mie.2019.06.005",
language = "English (US)",
isbn = "9780128183595",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "77--86",
editor = "Jungsan Sohn",
booktitle = "DNA Sensors and Inflammasomes",
}