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URL: https://dx.doi.org/10.17504/protocols.io.48qgzvw

Authors: Anika Wiegard, Christin Köbler, Katsuaki Oyama, Anja K. Dörrich, Chihiro Azai, Kazuki Terauchi, Annegret Wilde, Ilka Maria Axmann

Group: Axmann Lab, CyanoWorld

Summary: This protocol describes how to detect synthesis of [α32P]ATP from [α32P]ATP by recombinant KaiC proteins. Radioactive nucleotides are separated via thin layer chromatography using TLC PEI Cellulose F plates as stationary phase and LiCl as soluble phase. The principle of this method is based on Egli et al. (Egli M, Mori T, Pattanayek R, Xu Y, Qin X, Johnson CH.2012. Dephosphorylation of the core clock protein KaiC in the cyanobacterial KaiABC circadian oscillator proceeds via an ATP synthase mechanism. Biochemistry 51:1547-58.)

Associated Publications: Wiegard A, Köbler C, Oyama K, Dörrich AK, Azai C, Terauchi K, Wilde A, Axmann IM, Array. Journal of Bacteriology 202(4). doi: 10.1128/JB.00478-19

Affiliations: Institute for Synthetic Microbiology, Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University Duesseldorf, 40225 Duesseldorf, Germany, Institute of Biology III, Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany, Graduate School of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan, Institute for Microbiology and Molecular Biology, Justus-Liebig University, 35392 Giessen, Germany, College of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan; Graduate School of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan, College of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan; Graduate School of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan, Institute of Biology III, Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany, Institute for Synthetic Microbiology, Cluster of Excellence on Plant Sciences (CEPLAS), Heinrich Heine University Duesseldorf, 40225 Duesseldorf, Germany

External URL: https://doi.org/10.1128/JB.00478-19

Version: 1

Publication Date: 2019

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