Substrate Independent ATPase Activity May Complicate High Throughput Screening.
journal contribution
posted on 2017-12-16, 00:00 authored by Tuntland ML, Fung LW-MInorganic 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
Funding
The work was supported, in part, by a grant from the National Institutes of Health (U19 AI-56575).
History
Publisher Statement
This is the author’s version of a work that was accepted for publication in Analytical Biochemistry. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Analytical Biochemistry, 2016. 510: 18-20. DOI: 10.1016/j.ab.2016.07.016.Publisher
Elsevier Massonissn
0003-2697Issue date
2016-07-01Usage metrics
Categories
No categories selectedLicence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC