Fig. 4. Pyrophosphorylation of myoglobin-D127pS and subsequent characterization. (a) Pyrophosphorylation and refolding protocol for Myo-D127pS. Condition: 306 mM ZnCl2, 61.2 mM reagent 1, 2, or 3, and 49 μM protein. (b) CD spectra of myoglobin before (native) and after the pyrophosphorylation reaction (pyrophos.) according to (a) with reagent 1. (c) ESI-MS of Myo-D127pS pyrophosphorylated with reagent 1 for 100 min and refolded. (d) ESI-MS of Myo-D127pS pyrophosphorylated with reagent 3 for 180 min and refolded. Mass error of 4 ppm; mass resolution of 0.2 Da.
