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. 2020 Jun 4;16(6):e9355. doi: 10.15252/msb.20199355

Figure EV3. The metabolic compensation property of the Kai oscillator.

Figure EV3

  1. Fluorescence polarization measurements of oscillatory reactions at various [KaiA] and %ATP are fit to curves of the form FP (t) = A cos (2πT −1 t + ϕ) + bt + c to extract the normalized amplitude (100A/c; dimensionless) of the oscillator; see Fig 2D for the periods (T) of the same reactions. Reactions with an amplitude A < 0.5 are considered to be non‐oscillatory.
  2. Representative traces demonstrating the effect of %ATP at 1.25 μM KaiA; the polarization data are shifted vertically to avoid overlaps and horizontally to align the first peaks.
  3. Fluorescence polarization measurements of oscillatory reactions at three %ATP conditions in the presence of 0.0, 2.5, and 4.5 μM KaiC S431A/T431A (AA). The period (left) and amplitude (right) of the reactions are extracted using the same curve fit method as for panel A; non‐oscillatory reactions are not shown.
  4. SDS–PAGE gel image of the supernatant from the KaiB‐FLAG immunoprecipitation experiment.
  5. Simulated KaiC AA titration experiment using the modified Phong model. The points are model predictions, and the solid line is a linear fit. The dotted line is the linear fit to the experimental results (see Fig 3G). The simulations are carried out at 100% ATP, 1.5 μM KaiA, and K D = 10−3 μM condition.
  6. Comparison of the metabolic compensation property of the Phong model without (left) or with (right) a phosphorylation threshold at K D = 10−3 μM. The model exhibits phase decoherence at low %ATP without a phosphorylation threshold.