Fig. 1: Identification of the CaM-binding site in PYK2.
From: PYK2 senses calcium through a disordered dimerization and calmodulin-binding element
a Structural domain arrangement of FAK and PYK2. The FERM, kinase and FAT domains are colour-coded. Flexible linker regions are shown in white. Tyrosines important for kinase activation through phosphorylation are indicated in red. Proline-rich (PR) motifs 1–3 are marked in grey. Previously suggested Ca2+/CaM-binding sites in PYK2 are shown in yellow, and the site determined in this study is in orange. b Representative immunoblot of input fractions of the indicated GFP-tagged PYK2 constructs immunolabelled with GFP and tubulin antibodies. c Top: Representative GFP immunoblot of GFP-PYK2 constructs associated with the CaM Sepharose beads labelled as in b. Bottom: Graphical representation of GFP immunoblot densitometry associated with beads normalised by corresponding input, presented as foldchange using PYK2 WT Ca2+ bead fractions as a reference. Bars correspond to the mean of 4–7 independent experiments, ±SEM. d CaM Sepharose bead assay using recombinant purified FERM domains of PYK2 and FAK (left) and KFL regions of PYK2 and FAK (right). For each construct the input (inp.) and the bead fractions were run in adjacent lanes as indicated.
