Figure 3.

KIN-29 is a functional kinase. 293T cells were transfected with C-terminal flag tagged kin-29 constructs (top panel). KIN-29-KU contains amino acids 1–354, which includes the kinase domain and UBA domain. KIN-29K contains amino acids 1–300 which includes only the kinase domain and KIN-29K(K45R) contains amino acids 1–300 with a point mutation at position 45 that changes a lysine to an arginine. FLAG-tagged proteins were immunoprecipitated using anti-Flag antibody and in vitro kinase assays performed. Full length KIN-29 is capable of autophosphorylation similar to mammalian Tβ RII (TGFβ type II RSK) (top panel). Truncating the C-terminal domain of KIN-29 prevents autophosphorylation.