Table 1.
Binding of TPI to IOV
| Sample | Hemolysate, gHb/liter | TPI added, milliunits | TPI bound
|
||
|---|---|---|---|---|---|
| Percent | milliunits/mg IOV | ng/mg IOV | |||
| Propositus | 62.7 | 189 | 3.27 | 30.93 | 306.3 |
| 441 | 2.54 | 55.85 | 558.5 | ||
| Brother | 60.4 | 346 | 4.24 | 72.94 | 486.0 |
| 806 | 4.26 | 172.6 | 1016 | ||
| Control | 62.8 | 17,400 | 0.66 | 576.6 | 57.7 |
| 40,600 | 0.50 | 1,015.2 | 101.5 | ||
Binding experiments were carried out as described in Materials and Methods. A total of 200 μg of IOV from the normal and deficient cells were incubated with 300 or 700 μl of hemolysates from the same sources at 25°C for 30 min. The total protein contents of the hemolysates were similar. The estimation of the amount of the bound enzyme was based on the measured activities by using specific activities of 300 units/mg, 450 units/mg, and 9,800 units/mg for propositus, brother, and normal control TPIs, respectively. “Percent” represents the bound fraction of added TPI. The standard error of the determinations was ± 15% (n = 4). Data are from a typical set of experiments.