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GTP Cyclohydrolase 1 Activity Assays

TY Tao Yan
SG Shun Guo
TZ Tian Zhang
ZZ Zhimin Zhang
AL An Liu
SZ Song Zhang
YX Yuan Xu
YQ Yuhong Qi
WZ Weihe Zhao
QW Qinhui Wang
LS Lei Shi
LL Linna Liu
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Gch1 activity was determined as described previously (Nar et al., 1995). Briefly, the formation of dihydroneopterin triphosphate and the product of Gch1 were analyzed. The fused or free Gch1 protein was incubated in the dark with GTP assay buffer. Oxidation of dihydroneopterin triphosphate was performed with hydrogen chloride and potassium iodide/iodine in the dark. Samples were decolorized with ascorbic acid, vortexed, neutralized with sodium hydroxide and dephosphorylated with alkaline phosphatase. Samples were then centrifuged, and neopterin was quantified with an enzyme-labeled instrument (TECAN Infinite M200 Pro, Männedorf) with fluorescence detection.

Copyright and License information:  ©2021 Yan, Guo, Zhang, Zhang, Liu, Zhang, Xu, Qi, Zhao, Wang, Shi and Liu.
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