Table 1.
Kinetic analysis of the 2H, 13C labeled substrates with WT SLO-1.a
| Linoleic acid | kcat (s−1) | KM (μM) |
|---|---|---|
| [2H31]-1b | 5.7 (0.1) | 7.8 (0.5) |
| 11,11-[2H2]-1c | 4.5 (0.2) | 7.7 (1.3) |
| 10-[13C]-11,11-[2H2]-1d | 4.7 (0.2) | 7.8 (1.0) |
| 10,11-13C2-11,11-[2H2]-1d | 4.5 (0.2) | 7.6 (1.0) |
a
Reactions were conducted with 3.5 nM WT SLO, prepared as previously described,5 at 30°C in 0.1 M borate, pH 9.0 buffer. Substrates were prepared at 1 mM working stocks in 0.1 M sodium borate (pH 9.0) buffer. Prior to use, the concentration of the effective substrate was confirmed enzymatically. Each kinetic data set was conducted with 7 substrate concentrations. For each substrate analyzed, the resulting kinetic parameters are averages from 3 individual data sets.
b
Substrate is commercially available from Cambridge Isotopes. The first order rate (kcat) is from ref 22
c
Synthesized from our lab previously; kcat is consistent with previous report20.
d
Synthesized here.