Table 1.
Activity of T. brucei AdoMetDC and AdoMetDC/prozyme complexes.
|
TbAdoMetDC proteins |
Prozyme proteins |
kcat/Km (s−1M−1) |
|||
|---|---|---|---|---|---|
|
TbAdoMetDC monomer |
TbAdoMetDC/prozyme heterodimer |
||||
(+) Put |
(−) Put |
(+) Put |
(−) Put |
||
Wild-type |
Wild-type |
9.7 ± 3.5 |
0.48 ± 0.08 |
3.2 ± 0.6×103 |
2.6 ± 0.3×103 |
Δ16 |
Wild-type |
16±3* |
16±11* |
28±5* |
18±3* |
Δ26 |
Wild-type |
0.32 ± 0.07 |
0.11 ± 0.02 |
7.6 ± 1.7 |
2.2 ± 0.3 |
H172A |
Wild-type |
ND |
ND |
2.1 ± 0.4×103 |
93 ± 42 |
W137A/M146A |
Wild-type |
ND |
ND |
87 ± 8 |
5.0 ± 0.9 |
Wild-type |
M148'A/Y152'A |
as above |
as above |
0.66 ± 0.23×103 |
16 ± 4 |
* data taken from (Velez et al., 2013). Data were collected in the presence of 4 mM putrescine (Put), except for the Δ16 mutant (5 mM putrescine*). In all cases, the heterodimer with the wild-type counterpart formed with sufficient affinity that the subunits could be copurified as a stable complex. Error represents the standard deviation for the fit of triplicate data points. ND, not determined.