Specific Detection of Cellular Glutamine Hydrolysis in Live Cells Using HNCO Triple Resonance NMR

Title
Specific Detection of Cellular Glutamine Hydrolysis in Live Cells Using HNCO Triple Resonance NMR
Authors
차진욱이수진He Wen박성혁
Issue Date
2016-11
Publisher
ACS Chemical Biology
Citation
VOL 11, NO 11-3145
Abstract
Glutamine plays key roles as a biosynthetic precursor or an energy source in cancers, and interest in its metabolism is rapidly growing. However, the proper evaluation of glutamine hydrolysis, the very first reaction in the entire glutaminolysis, has been difficult. Here, we report a triple resonance NMR-based assay for specific detection of glutaminase activity carrying out this reaction using stable-isotope labeled glutamine. Compared to conventional methods involving coupled enzyme assays, the proposed approach is direct because it detects the presence of the H– N– CO amide spin system. In addition, the method is unique in enabling the measurement of glutamine hydrolysis reaction in real-time in live cells. The approach was applied to investigating the effects of a glutaminase inhibitor and the inhibitory effects of glucose on glutamine metabolism in live cells. It can be easily applied to studying other signals that affect cellular glutamine metabolism
URI
http://pubs.kist.re.kr/handle/201004/64890
ISSN
1554-8929
Appears in Collections:
KIST Publication > Article
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