Figure 3.

(A) Expression and purification of His-NEK1 kinase domain (NEK1∆CT). (B) NEK1∆CT was catalytically active and ATP/ADP conversion (kinase activity) was linear with the enzyme amount. (C) In vitro phosphorylation reactions of YAP using His-NEK1 and MK5 kinases in presence of [γ-³²P] ATP. (D) In vitro phosphorylation of YAP using His-NEK1 and MK5 kinases for preparative isolation for MS determination of phosphopeptides. (E) His-NEK1 also phosphorylated YAP on Tyr, as demonstrated by immunoreactivity with pY antibody.