Table 2.
Requirements of the Glu-AdT reaction
| Enzyme/substrate combination* | pmol of glutamine recovered |
|---|---|
| Glu-AdT + Glu-tRNAGln + ATP + Gln + MgCl2 | 10.0 |
| Glu-AdT + Glu-tRNAGln + ATP + Asn + MgCl2 | 3.8 |
| Glu-AdT + Glu-tRNAGln + ATP + NH4Cl + MgCl2 | 2.0 |
| Glu-AdT + Glu-tRNAGlu + ATP + Gln + MgCl2 | <0.02† |
| Glu-AdT + Glu-tRNAGln + ATP + Gln | <0.02 |
| Glu-AdT + Glu-tRNAGln + CTP + Gln + MgCl2 | <0.02 |
| Glu-AdT + Glu-tRNAGln + GTP + Gln + MgCl2 | <0.02 |
| Glu-AdT + Glu-tRNAGln + UTP + Gln + MgCl2 | <0.02 |
| Glu-AdT + Glu-tRNAGln + ATP + Gln + MgCl2 + Ab‡ | <0.02 |
*
The assay conditions are described in Materials and Methods.
†
The amount of [14C]glutamine recovered was below the 0.02 pmol limit of detection of the assay protocol.
‡
Ab, polyclonal antibodies raised against the heterotrimeric complex.