Fig. 2: DYRK2 specifically phosphorylates the MYO6 S267 peptide and the MYO6 full-length protein. | Nature Communications

Fig. 2: DYRK2 specifically phosphorylates the MYO6 S267 peptide and the MYO6 full-length protein.

From: Motor ___domain phosphorylation increases nucleotide exchange and turns MYO6 into a faster and stronger motor

Fig. 2

a Activities (HotSpotTM kinase assay) of 31 kinases with respect to the MYO6 peptide, compared with their ideal substrates. b A representative SDS-PAGE image of recombinant DYRK2 purified from E. coli. The purification has been performed several times. c Schematic overview of the ADP-glow assay used to measure DYRK2 activity. d ADP-glowTM kinase assays show that recombinant DYRK2 has ATPase activity with respect to both DYRK-specific substrate and wild-type MYO6 peptide containing the SSP and the ASP sequence, but not to peptides with the mutant SAP or AAP sequence. e DYRK2 shows ATPase activity towards the MYO6 motor ___domain in vitro. Before the ADP-glow assay full length MYO6 was heat-treated at 37 °C or 60 °C to inactivate endogenous ATPase activity that would interfere with the kinase assay. f MYO6 was immunoprecipitated from A431 cells grown in serum-free media (starved) and incubated with purified DYRK2 for 1 h before determining the level of S267 phosphorylation by mass spectrometry. g DYRK2 does not phosphorylate MYO6 at T405 using the wildtype MYO6 T405 peptide in the ADP-glow assay in vitro.

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