Table 2.

Binding affinities of mono- and dinucleotide probes for DcpS together with fluorescence enhancement factors (g).

L.p	Protein	KD / nM	g = FLbound / FLfree
1d	DcpS	137 ± 5	0.91 ± 0.01
1e	DcpS	105 ± 12	0.88 ± 0.01
1f	DcpS	58 ± 2	0.80 ± 0.01
2e	DcpS	152 ± 32	0.93 ± 0.10
2f	DcpS	44 ± 5	0.71 ± 0.03
2g	DcpS	53 ± 13	1.88 ± 0.12
2h	DcpS	18 ± 6	1.54 ± 0.04
2i	DcpS	12.6 ± 0.9	1.12 ± 0.01

FA experiments were performed in black 96-well plates using a Biotek Synergy H1 plate reader. Each well (200 µL) contained a carboxyfluorescein-tagged probe (0.5, 1, 2, or 10 nM) and increasing concentrations of the desired protein (from 0 to 2.5 µM).