Background: Slik kinase has catalytic activity-dependent and -independent functions. Results: Mutation of activation segment phosphorylation sites abolishes both catalytic and non-catalytic activities; non-catalytic function also depends upon localization via the C-terminal domain. Conclusion: Slik is regulated by both localization and phosphorylation. Significance: Conformational activation can control not only catalytic but also non-catalytic activities of kinases.