Table 1 Binding affinities of HaloTag-635-LIG3α-XRCC1-YFP with nicked λ DNA substrates
From: The zinc finger of DNA ligase 3α binds to nucleosomes via an arginine anchor
PROTEIN | DNA (10 NICKED λ DNA) | LIFETIMES (s) | koff (s–1) | N | kon app (s-1) | N’ | kon (M–1s–1) | KD (nM) |
|---|---|---|---|---|---|---|---|---|
Halo-LIG3α | 3’OH 5’P | 5.1 ± 0.02 | 0.2 ± 0.0007 | 2078 | 0.1 ± 0.004 | 256 | 1.7 ± 0.04 ×109 | 0.12 ± 0.003 |
YFP-XRCC1 | 3’OH 5’P | 6.3 ± 0.03 | 0.2 ± 0.0007 | 1219 | 0.1 ± 0.004 | 140 | 1.1 ± 0.03 ×108 | 1.5 ± 0.04 |
Halo-LIG3α | 3’OH 5’OH | 3.6 ± 0.04 | 0.3 ± 0.003 | 701 | 0.06 ± 0.01 | 359 | 1.6 ± 0.3 ×108 | 1.6 ± 0.3 |
YFP-XRCC1 | 3’OH 5’OH | 4.7 ± 0.07 | 0.2 ± 0.003 | 289 | 0.04 ± 0.002 | 167 | 1.3 ± 0.06 ×107 | 17 ± 0.9 |
Halo-Del-ZnF- LIG3α | 3’OH 5’P | 13 ± 0.3 | 0.08 ± 0.002 | 119 | 0.04 ± 0.003 | 62 | 8.6 ± 0.6 ×108 | 0.93 ± 0.007 |
YFP-XRCC1 | 3’OH 5’P | 15 ± 0.4 | 0.06 ± 0.002 | 102 | 0.005 ± 0.003 | 29 | 4.2 ± 3 ×107 | 1.6 ± 0.9 |
Halo-ZnF-BRCT-LIG3α | 3’OH 5’P | 13 ± 0.3 | 0.08 ± 0.002 | 110 | 0.02 ± 0.001 | 26 | 2.4 ± 0.1 ×107 | 3.2 ± 0.2 |
YFP-XRCC1 | 3’OH 5’P | 16 ± 0.3 | 0.06 ± 0.001 | 80 | 0.02 ± 0.004 | 21 | 1.6 ± 0.3 ×107 | 4.0 ± 0.8 |
Halo-K421A-LIG3α | 3’OH 5’P | 1.7 ± 0.02 | 0.6 ± 0.007 | 654 | 0.05 ± 0.002 | 52 | 1.5 ± 0.06 ×108 | 0.40 ± 0.01 |
YFP-XRCC1 | 3’OH 5’P | 1.4 ± 0.02 | 0.7 ± 0.01 | 185 | 0.06 ± 0.006 | 11 | 1.7 ± 0.2 ×108 | 4.1 ± 0.4 |
