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. 2001 Mar 16;2:2. doi: 10.1186/1471-2091-2-2

Figure 2.

Figure 2

In vivo phosphorylation of DokA during osmotic stress. PHKR was immunoprecipitated from the cell lysate of radioactively labeled and osmotically shocked Dictyostelium PHKR cells. Purified proteins were separated by SDS-PAGE, blotted onto a PVDF membrane and subjected to autoradiography. Panel A: PHKR is phosphorylated in vivo during osmotic shock (t = 20 min, 40 min). Mutation of the conserved histidine in PHKR does not affect the phosphorylation of PHKR. Before the osmotic shock (t = 0 min), neither of the two DokA fragments are phosphorylated. The radioactive label can not be removed by treating the membrane with 1 N HCl for 1 h. It was shown by immunostaining with the mAb 426/HK antibody that equal amounts of protein were purified at each point in time. Panel B: PHKR precipitated from untreated cells is not radioactively labeled.