ArticleMethods in enzymology2015
Detecting KaiC phosphorylation rhythms of the cyanobacterial circadian oscillator in vitro and in vivo.
Article in Methods in enzymology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Effects of sodium chloride on circadian period and temperature compensation of KaiC phosphorylation.Scientific reports · 2026Article
- Damped Oscillating Phosphoryl Transfer Reaction in the Cyanobacterial Circadian Clock.ACS omega · 2023Article
- Shift in Conformational Equilibrium Underlies the Oscillatory Phosphoryl Transfer Reaction in the Circadian Clock.Life (Basel, Switzerland) · 2021Article
- The Circadian Clock-A Molecular Tool for Survival in Cyanobacteria.Life (Basel, Switzerland) · 2020Review
- CikA, an Input Pathway Component, Senses the Oxidized Quinone Signal to Generate Phase Delays in the Cyanobacterial Circadian Clock.Journal of biological rhythms · 2020Article
- Damped circadian oscillation in the absence of KaiA in Synechococcus.Nature communications · 2020Article
- For Whom the Clock Ticks: Clinical Chronobiology for Infectious Diseases.Frontiers in immunology · 2020Review
- A Combined Computational and Genetic Approach Uncovers Network Interactions of the Cyanobacterial Circadian Clock.Journal of bacteriology · 2016Article
- A dynamic interaction process between KaiA and KaiC is critical to the cyanobacterial circadian oscillator.Scientific reports · 2016Article
- Cross-talk and regulatory interactions between the essential response regulator RpaB and cyanobacterial circadian clock output.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
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Authors and funding
4 authors.
Funding
Abstract
The central oscillator of the cyanobacterial circadian clock is unique in the biochemical simplicity of its components and the robustness of the oscillation. The oscillator is composed of three cyanobacterial proteins: KaiA, KaiB, and KaiC. If very pure preparations of these three proteins are mixed in a test tube in the right proportions and with ATP and MgCl2, the phosphorylation states of KaiC will oscillate with a circadian period, and these states can be analyzed simply by SDS-PAGE. The purity of the proteins is critical for obtaining robust oscillation. Contaminating proteases will destroy oscillation by degradation of Kai proteins, and ATPases will attenuate robustness by consumption of ATP. Here, we provide a detailed protocol to obtain pure recombinant proteins from Escherichia coli to construct a robust cyanobacterial circadian oscillator in vitro. In addition, we present a protocol that facilitates analysis of phosphorylation states of KaiC and other phosphorylated proteins from in vivo samples.
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