TABLE 1.
Pulse-chase experiments for the trapping of 14CH3 groups with CODH/ACS or with ACS with 1 mm TiCit, 1 mm CoA, and 1.0 atm CO
The background of radioactivity detection was measured using a control without CODH/ACS or ACS. It was estimated as 1.2 μm. The concentrations given are final concentrations.
|
Time delay |
[CODH/ACS] |
[14CH3-CFeSP] pulse |
[CH3-CFeSP] chase |
[14C2-acetyl-CoA] formeda |
Percentage recovery in pulse/chaseb |
Expected percentage recoveryc |
|---|---|---|---|---|---|---|
| s | μm | μm | μm | μm | % | % |
| 100 | 50 | 6.2 | 0 | 3.80 ± 0.5 | ||
| 80 | 1.80 ± 0.5 | 47 ± 18 | 7 | |||
| 15 | 0 | 3.1 ± 0.5 | ||||
| 80 | 3.5 ± 0.5 | 110 ± 24 | 16 | |||
| 10 | 0 | 5.0 ± 0.5 | ||||
| 80 |
6.8 ± 0.5 |
135 ± 17 |
11 |
|||
|
[ACS] |
||||||
| s | μm | |||||
| 120 | 24 | 24 | 0 | 4.8 ± 0.5 | ||
| 70 |
5.7 ± 0.5 |
120 ± 16 |
26 |
a
Recovery was based on 14C recovered in acetyl-CoA as calculated by Equation 1
b
Percentage recovery was calculated by Equation 2
c
The expected percentage recovery is based on the isotope dilution if the methyl group must dissociate before product can be formed, equal to the ratio of [14CH3-CFeSP] to total [CH3-CFeSP]