Table 2. Elution volumes, 260/280 ratio, experimental and theoretical molecular weights of RPA, Tim-Tipin (TTP) and Tim-Tipin-RPA in presence of ssDNA substrates.
DNA substrate/ Protein:ssDNA [μM] | Protein mixture/ DNA substrate | Elution volume* [ml] | 260/280* | ExperimentalSEC# molecular weight [kDa] | Theoretical§ molecular weight [kDa] |
---|---|---|---|---|---|
RPA | 1.63 ± 0.02 | 0.63 ± 0.02 | 110 | 96.2 | |
Tim-Tipin | 1.46 ± 0.01 | 0.55 ± 0.01 | 321 | 162.3 | |
Tim-Tipin-RPA | 1.43 ± 0.02 | 0.56 ± 0.01 | 388 | 258.5 | |
60 nt | RPA | 1.54 ± 0.00 | 1.57 ± 0.05 | 194 | 114.6 |
16:32 | Tim-Tipin | 1.44 ± 0.01 | 0.98 ± 0.14 | 364 | 180.7 |
Tim-Tipin-RPA | 1.39 ± 0.01 | 1.20 ± 0.04 | 498 | 276.9 | |
60 nt | RPA | 1.38 ± 0.01 | 1.32 ± 0.03 | 531 | 403.2 |
32:8 | Tim-Tipin | 1.47 ± 0.01 | 0.74 ± 0.06 | 301 | 180.7 |
Tim-Tipin-RPA | 1.44 ± 0.01 (TTP) | 0.66 ± 0.05 | 364 | ||
1.33 ± 0.05 (RPA) | 1.13 ± 0.04 | 726 | |||
31 nt | RPA | 1.55 ± 0.03 | 1.46 ± 0.01 | 182 | 105.7 |
16:32 | Tim-Tipin | 1.46 ± 0.02 | 0.74 ± 0.01 | 321 | 171.8 |
Tim-Tipin-RPA | 1.40 ± 0.03 | 1.11 ± 0.04 | 468 | 268.0 | |
31 nt | RPA | 1.48 ± 0.02 | 1.28 ± 0.05 | 283 | 201.9 |
16:8 | Tim-Tipin | 1.47 ± 0.01 | 0.57 ± 0.02 | 301 | 171.8 |
Tim-Tipin-RPA | 1.41 ± 0.03 (TTP) | 0.87 ± 0.0 | 439 | ||
1.38 ± 0.03 (RPA) | 0.98 ± 0.01 | 531 | |||
14 nt | RPA | 1.63 ± 0.0 | 1.47 ± 0.24 | 110 | 100.5 |
16:32 | Tim-Tipin | 1.48 ± 0.01 | 0.54 ± 0.02 | 283 | 166.6 |
Tim-Tipin-RPA | 1.45 ± 0.02 (TTP) | 0.66 ± 0.08 | 342 | ||
1.62 ± 0.01 (RPA) | 1.27 ± 0.05 | 117 | |||
14 nt | RPA | 1.63 ± 0.0 | 1.27 ± 0.06 | 110 | 100.5 |
16:8 | Tim-Tipin | 1.48 ± 0.01 | 0.54 ± 0.0 | 283 | 166.6 |
Tim-Tipin-RPA | 1.45 ± 0.02 (TTP) | 0.64 ± 0.05 | 342 | ||
1.62 ± 0.01 (RPA) | 1.22 ± 0.04 | 117 | |||
ssDNA only | 60 nt | 1.70 | 1.82 | 64 | 18.4 |
0:16 | 31 nt | 1.70 | 1.76 | 64 | 9.5 |
14 nt | 1.88 | 2.34 | 21 | 4.3 | |
Thyroglobulin | 1.32 ± 0.02* | 670 | |||
γ-globulin | 1.63 ± 0.03* | 158 | |||
Ovalbumin | 1.76 ± 0.01* | 44 | |||
Myoglobin | 1.91 ± 0.02* | 17 | |||
Vitamin B12 | 2.15 ± 0.02* | 1.35 |
#Molecular weights estimated based on the comparison with molecular mass standard proteins.
§Molecular weights calculated based on the amino acid sequence.
* Average numbers and standard deviations of at least two independent measurements are shown.