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. 2019 Dec 4;20(24):6120. doi: 10.3390/ijms20246120

Figure 13.

Figure 13

AtCRK5 phosphorylates the PIN1 hydrophilic loop in vitro. We performed in vitro radioactive phosphorylation assay with His6-AtCRK5 and two substrates: Myelin Basic Protein (MBP) as a positive control, and GST-PIN1-HL loop. White asterisks indicate the phosphorylation event on MBP (* asterisk) and GST-PIN1HLloop (** asterisk) proteins, respectively. We carried out glutathione S-Transferase (GST) column purification after the kinase reactions shown in the last three columns to remove the HIS6-CRK5 kinase because the His6-CRK5 and the GST-PIN1 protein sizes are nearly identical and we could not distinguish the phosphorylation signals.