Table 2. Immobilized [3H]acetylated N-acetyltransferase-His8Flag intermediate cannot be detected in immunoprecipitation experiments with anti-Flag M2 beads.
| With lipids1 at pH 6.5, 60 min | Immobilized on Flag-Beads | Unbound & washes | ||
| Total cpm | Activity2 | Total cpm | Activity | |
| Transfected1 | 1,2303 ±40 | 232 (13) | 87,300±2,100 | 1.2 |
| Untransfected1 | 1,240 ±60 | 1.1 | 85,000±2,100 | ND4 |
1
Anti-Flag beads were incubated with extracts from HeLa cell expressing (Transfected) or not expressing (Untransfected) N-acetyltransferase-His8Flag, washed and then incubated with [3H]acetyl-CoA for the given time and pH, at room temperature and with or without 1.3 mM of lipids containing 20% PI.
2
N-acetyltransferase activity in nmoles of MU produced/hour, and the concentration of protein (pmoles), calculated based on the specific activity of the purified transferase, are given.
3
The data represent three independent beads binding experiments and assays.
4
Not detectable.