Table 2.

Phe-tRNA^Phe variant parameters^a

	E. coli	E. coli Cy3	E. coli Cy5	E. coli Cy5.5	Yeast	Yeast Cy3	Yeast Cy5
Phe-tRNAPhe preparation (%)
Labeling		80	90	79		92	93
Charging	28	25	15	26	44	37	30
Empirical parameters
tLb (2 µM)	1.00 ± 0.05	1.47 ± 0.11	1.13 ± 0.22	1.13 ± 0.08	1.20 ± 0.11	1.27 ± 0.11	2.33 ± 0.15
tLb (4 µM)	1.27 ± 0.11	1.67 ± 0.04	1.07 ± 0.08	NA	1.07 ± 0.07	1.18 ± 0.11	2.27 ± 0.25
S1c (2 µM)	1.00 ± 0.22	0.48 ± 0.17	0.42 ± 0.04	0.71 ± 0.19	0.79 ± 0.04	0.42 ± 0.02	0.21 ± 0.02
S1c (4 µM)	2.34 ± 0.06	0.60 ± 0.07	0.43 ± 0.04	NA	1.43 ± 0.05	0.50 ± 0.03	0.09 ± 0.01
A1d (2 µM)	1.00 ± 0.09	0.87 ± 0.10	0.35 ± 0.05	0.55 ± 0.17	1.02 ± 0.04	0.68 ± 0.01	0.38 ± 0.02
A1d (4 µM)	2.89 ± 0.06	1.52 ± 0.09	0.81 ± 0.03	NA	2.41 ± 0.01	1.24 ± 0.06	0.50 ± 0.02
S2e (2 µM)	1.00 ± 0.14	1.17 ± 0.15	0.63 ± 0.07	0.37 ± 0.18	1.14 ± 0.08	0.71 ± 0.09	0.68 ± 0.04
S2e (4 µM)	2.61 ± 0.12	1.54 ± 0.12	0.78 ± 0.11	NA ±	1.74 ± 0.03	1.03 ± 0.05	0.73 ± 0.09
Relative 14C-Phe incorporationf
	1.00 ± 0.02	0.98 ± 0.02	0.13 ± 0.01	NA	0.66 ± 0.01	0.43 ± 0.02	0.12 ± 0.01
Simulation
Relative	1g	0.1	0.025	0.2	1g	0.05	0.025
Phase 2 steady-state Phe-tRNAPhe concentration (µM)	0.1	0.1	0.01	0.01	0.15	0.025	0

^aData at 2 µM added Phe-tRNA^Phe are presented in Figure 4B and C. Parallel results obtained at 4 µM Phe-tRNA^Phe are summarized in the table. Experiments were performed using 2–5 repeats. Data obtained with 2 µM E. coli tRNA variants was repeated with two different CFPS batches. All values were normalized to 2 µM E. coli Phe-tRNA^Phe.

^bLag time.

^cPhase-1-slope.

^dPhase-1-amplitude.

^ePhase-2-slope.

^fAfter 30-min incubation.

^gEqual to k₂.