TABLE 1.
In vitro adenylation and in vivo activity of MccA heptapeptide point mutantsa
| Peptide sequence | Adenylation by MccB in vitro |
In vivo activity |
|
|---|---|---|---|
| Wild-type cells | ΔyejB cells | ||
| MRTGNAN | + | + | − |
| MRTGNAD | − | − | − |
| MRTGNAQ | − | − | − |
| MRAGNAN | −/+ | − | − |
| MRLGNAN | +/− | + | − |
| MRSGNAN | + | + | − |
| MRCGNAN | +/− | + | − |
| MRKGNAN | − | − | − |
| MR(d-Thr)GNAN | − | − | − |
| MRTWNAN | +/− | + | − |
| GRTGNAN | − | − | − |
| ARTGNAN | − | − | − |
| IRTGNAN | +/− | + | − |
| VRTGNAN | − | − | − |
| LRTGNAN | +/− | + | − |
| FRTGNAN | + | + | − |
| NRTGNAN | − | − | − |
| QRTGNAN | − | − | − |
| DRTGNAN | − | − | − |
| ERTGNAN | − | − | − |
| KRTGNAN | − | − | − |
a
The table presents the results of in vitro adenylation by MccB of each peptide listed as detected by MALDI-MS analysis of reactions conducted under standard conditions (see, for example, Fig. 2 and Materials and Methods) and the results of bioactivity testing of 10-μl aliquots of products of completed adenylation reactions on lawns of wild-type or yejB mutant E. coli cell lawns. +, a clear growth inhibition zone was detected; −, no growth inhibition zone was observed; +/−, incomplete conversion.