TABLE 1.
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 |
Recovery was based on 14C recovered in acetyl-CoA as calculated by Equation 1
Percentage recovery was calculated by Equation 2
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]