Table 1.
Substrates investigated in the phosphorylation reaction.
| Substrate | Potential products | Products confirmed by co-injection | Yield (%) |
|---|---|---|---|
| 5′ AMPa | ADPb, c | 5′ ADP | 5′ ADP: (2.0 ± 0.3) |
| ATPc | 5′ ATP | 5′ ATP: (0.4 ± 0.2) | |
| 5′ ADPa | AMPc | 5′ AMP | 5′ AMP: (68.8 ± 3.7) |
| ATPc | 5′ ATP | 5′ ATP: (1.1 ± 0.1) | |
| Aa | AMPc | 5′/3′/2′ AMP | 5′ AMP: (26.7 ± 6.7) |
| ADPc | 5′ ADP | 5′ ADP: (2.2 ± 0.1) | |
| ATP | Traces | ||
| Ga | GMPc | 5′ GMP | 5′ GMP: (7.1 ± 1.6) |
| GDPc | 5′ GDP | Traces | |
| GTP | Traces | ||
| Ca | CMPc | 5′ CMP | 5′ CMP: (23.2 ± 2.4) |
| CDPc | 5′ CDP | 5′ CDP: (3.0 ± 0.3) | |
| CTP | Traces | ||
| Ua | UMPc | 5′ UMP | 5′ UMP: (29.6 ± 2.5) |
| UDPc | 5′ UDP | 5′ UDP: (1.5 ± 0.0) | |
| UTP | Traces | ||
| dAa | dAMP | Traces | |
| dGa | dGMP | Traces | |
| dCa | dCMPc | 5′ dCMP | 5′ dCMP: (32.6 ± 5.5) |
| dCDPc | 5′ dCDP | n. d.d | |
| dCTPc | 5′ dCTP | Traces | |
| dTa | dTMPc | 5′ dTMP | 5′ dTMP: (31.7 ± 9.4) |
| dTDPc | 5′ dTDP | 5′ dTDP: (3.6 ± 0.2) | |
| dTTPc | 5′ dTTP | Traces | |
| GL | GL phosphatee | GL-1-phosphate, GL-2-phosphate | n. d.c |
| Cyclic GL phosphate | n. d.c | ||
| GL phosphonate | n. d.c | ||
| GL2 phosphate | n. d.c | ||
| GL2 diphosphate | n. d.c | ||
| GL2 triphosphate | n. d.c | ||
| GL3 triphosphate | n. d.c | ||
| Mixed (non-)cyclic GL diphosphate | n. d.c | ||
| Mixed GL2 (non-)cyclic diphosphate | n. d.c | ||
| GA | GA phosphate | n. d.c | |
| GA phosphonate | n. d.c | ||
| GA2 phosphonate | n. d.c | ||
| Rib | Rib phosphatee | Rib-5-phosphate | n. d.c |
| Sodium Lc | Lc phosphate | n. d.c | |
| Cyclic Lc phosphate | n. d.c | ||
| Lc phosphonate | n. d.c | ||
| Lc phosphate phosphonate | n. d.c | ||
| Lc triphosphonate | n. d.c | ||
| Lc2 phosphate | n. d.c | ||
| Ser | Ser phosphatee | O-phospho Ser | n. d.c |
| Ser phosphonate | n. d.c | ||
| Ser diphosphate | n. d.c | ||
| Ser2 phosphate | n. d.c | ||
| Ser2 diphosphate | n. d.c | ||
| Ser triphosphate | n. d.c | ||
| Ser triphosphonate | n. d.c |
Substrates (100 mM, 1.0 eq.) were phosphorylated with H3PO3 (3.0 eq.) in the presence of urea (1.0 eq.) for 7 d. Yield errors refer to ± s.d. and were obtained by double determination via CE.
aComplete product mixture is displayed in Supplementary Data 1.
bFor bold labelled products, at least one signal was assigned to a particular isomer by co-injection with a reference.
cCE measurement with co-injection.
dNot determined.
eCE-MS measurement with co-injection.