Table 1.
N-terminal Modification of Reporter Peptide After Single Addition of Adenosyl Donor
| Donor | Additivesa | Yield (%)b |
|---|---|---|
| 4a (Phe) | 92.7 ± 5.2 | |
| Phe-pdCpA | 95.2 ± 4.3 | |
| 4a (Phe) | 1 mM ATP | 86.1 ± 0.8 |
| 4a (Phe) | 5 mM ATP | 79.1 ± 0.4 |
| 4a (Phe) | 5 mM AMP | 88.2 ± 0.8 |
| 4a (Phe) | 5 mM A | 93.1 ± 1.1 |
| 4a (Phe) | 1 mM pdCpA | 90.4 ± 0.9 |
| 4b (Leu) | 80.4 ± 0.9 | |
| 4c (Azf) | 78.5 ± 5.2 | |
| 4d (Nap) | 95.4 ± 0.4 | |
| 4d (Nap) | 100 μM PheLysAlaAcm | 93.8 ± 0.3 |
| 4e (Mef) | 0.6 ± 0.1 | |
| 4f (Acf) | 5f not observed | |
| 4g (N3f) | 5g not observedc | |
| 4h (Mcm) | 5h not observed | |
| 4i (Bzf) | 9.2 ± 0.3 |
a
Standard AaT-catalyzed transfer conditions with or without potential inhibitor added. See description under Experimental Procedures.
b
Yield determined by integration of HPLC chromatogram peak areas at 325 nm, described in Experimental Procedures. Standard deviations are reported for an average of at least 6 experiments with at least 2 different AaT preparations.
c
HPLC and MALDI MS analysis indicate that the observed product is 5a not 5g.