TABLE 2.
Kinetic parameters of 3D-RNA complex assembly and stability for FMDV 3Dwt, 3D(K18E), 3D(K20E), 3D(K18A), 3D(K20A), and 3D(KAKA)a
| Enzyme | kassembly (10−4 s−1)b |
koffc |
|
|---|---|---|---|
| ERn (10−3 s−1) | ERn+1 (10−4 s−1) | ||
| 3Dwt | 25 ± 4 | 28 ± 1 | 10 ± 4 |
| 3D(K18E) | 25 ± 5 | 60 ± 1 | 19 ± 4 |
| 3D(K20E) | 18 ± 7 | 67 ± 2 | 25 ± 3 |
| 3D(K18A) | 30 ± 5 | 34 ± 1 | 13 ± 2 |
| 3D(K20A) | 20 ± 3 | 42 ± 2 | 18 ± 2 |
| 3D(KAKA) | 26 ± 6 | 67 ± 2 | 20 ± 2 |
a
Kinetic parameters were obtained from the data shown in Fig. 2 according to the procedures detailed in Materials and Methods.
b
Rate of assembly of the different polymerases to sym/sub-U.
c
Rate of dissociation of the complex 3D-sym/sub-U either with no nucleotide incorporated (ERn) or with incorporation of AMP (ERn+1). ERn, enzyme-RNA complex, where n is the length of the primer.