Figure 1: ANKS6 interacts with NEK8, enhancing NEK8 kinase activity.
From: ANKS6 is the critical activator of NEK8 kinase in embryonic situs determination and organ patterning
(a) IP of FLAG-NEK8 from mIMCD3 cells stably transduced with a retroviral FLAG-NEK8 expression construct reveals a number of distinct co-precipitating bands on silver stain. ANKS6 is identified by mass spectrometry. (b) Mass spectrometric analysis of the FLAG-IP eluate reveals a number of distinct peptide fingerprints, among which ANKS6 was elected as an interesting candidate protein for further analysis. (c) ANKS6 co-precipitates with NEK8, is phosphorylated by NEK8 and promotes NEK8 autophosphorylation in an IP/kinase assay. No phosphorylation activity was observed in the absence of ANKS6, or in the setting of a K33M ‘kinase-dead’ allele. (d) ANKS6 enhances kinase activity of wild-type NEK8, as well as the pathogenic NEK8jck mutant, towards itself, the kinase, and towards α-casein, a nonspecific phosphorylation substrate.
