期刊
ANALYTICAL BIOCHEMISTRY
卷 510, 期 -, 页码 18-20出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2016.07.016
关键词
Malachite green; High throughput screening; Substrate-independent ATPase activity
资金
- National Institutes of Health [U19 AI-56575]
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. (C) 2016 Elsevier Inc. All rights reserved.
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