Extended Data Fig. 6: Comparisons between biochemically measured rubisco kinetic parameters and those same parameters as inferred from fitness values.
a and b) Fitness vs. kcat values, fitness error is the standard error of the mean for 9 replicates, c and d) \({\widetilde{K}}_{C}\) vs. KC values, \({\widetilde{K}}_{C}\) error bars reflect the inner quartiles of the bootstrap fits (see Methods). Measurements are from the literature in a and c, values are measured in this study by the spectrophotometric assay in b and d. Black points in b were purified 3 independent times (x-axis error bars are standard error), all other data in grey are from individual purifications and have no errors reported. Inset shows mutants with fitness values near or above 1 (WT-level). Dashed line indicates a 1:1 correspondence between fitness and in vitro measurements, WT is indicated with a square. X-axis error bars in a and c are taken from the literature when available. X-axis errors in d and Y-axis errors in a-d are explained in the methods. N = 3 biological replicates in all cases. Outlier mutation is labelled in a and b and is discussed in Methods. Red indicates \({\widetilde{K}}_{C}\) estimates with coefficient of variation >1. e) \({\widetilde{K}}_{C}\) coefficient of variation as a function of fitness. f) \({\widetilde{V}}_{\max }\) coefficient of variation as a function of \({\widetilde{V}}_{\max }\). g) \({\widetilde{K}}_{C}\) coefficient of variation as a function of fitness \({\widetilde{V}}_{\max }\) coefficient of variation. h) Correlation of \({\widetilde{V}}_{\max }\) and Fitness. Only mutants with a coefficient of variation <1 are plotted here; mutants with coefficients of variation >1 typically have low fitness and are thus harder to fit to a Michaelis-Menten model.
