Substrate independent ATPase activity may complicate high throughput screening.

Abstract

Inorganic phosphate release, [Pi], is often measured in an enzymatic reaction in a high throughput setting. Based on the published mechanism, we designed a protocol for our screening for inhibitors of SAICAR synthetase (PurC), and we found a gradual increase in [Pi] in positive control samples over the course of the day. Further investigation indicated that hydrolysis of ATP catalyzed by PurC, rather than substrate-related phosphate release, was responsible for a partial contribution to the signals in the control samples. Thus substrate-independent ATPase activity may complicate high throughput screening.

DOI: 10.1016/j.ab.2016.07.016

Cite this paper

@article{Tuntland2016SubstrateIA, title={Substrate independent ATPase activity may complicate high throughput screening.}, author={Micheal L Tuntland and Leslie W-M Fung}, journal={Analytical biochemistry}, year={2016}, volume={510}, pages={18-20} }