Fig. 4: PLK1 phosphotyrosine peptides and their in-vitro dephosphorylation by the EYA4 phosphatase domain. | Nature Communications

Fig. 4: PLK1 phosphotyrosine peptides and their in-vitro dephosphorylation by the EYA4 phosphatase domain.

From: The Eyes Absent family members EYA4 and EYA1 promote PLK1 activation and successful mitosis through tyrosine dephosphorylation

Fig. 4

A Schematic showing an example of immunopurified and Coomassie stained PLK1 which was in-gel digested and used in downstream pilot and PRM mass spec experiments. B PLK1 tyrosine phosphosites detected by mass spec with their positions designated on a PLK1 gene schematic. C The purified phosphatase domain of EYA4 (left, Coomassie-stained SDS-PAGE gel) was used in in-vitro phosphatase assays with synthetic phosphopeptides in duplicate. Peptides dephosphorylated with relatively fast kinetics (n = 2, pY445, Km 0.46 mM; pY425, Km 0.34 mM; and a positive control peptide from H2AX pY142, 0.51 mM) were fit to Michaelis-Menten curves (middle panel). Peptides with slower kinetics, which could not be fit to a Michaelis-Menten curve at the substrate concentrations tested, were fit using linear regressions (right panel). Individual datapoints represent means. Error bars represent standard deviations. Source data are provided as a Source data file.

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