The protein kinase D1 COOH terminus: marker or regulator of enzyme activity?

Protein kinase D1 (PKD1) is a Ser/Thr kinase implicated in a wide variety of cellular responses. PKD1 activation is generally attributed to a PKC-dependent pathway that leads to phosphorylation of the activation loop at Ser744/Ser748. This modification increases catalytic activity, including that toward an autophosphorylation site (Ser916) in a postsynaptic density-95/disks large/zonula occludens-1 (PDZ)-binding motif at the extreme COOH terminus. However, there is growing evidence that PKD1 activation can also result from a PKC-independent autocatalytic reaction at Ser744/Ser748 and that certain stimuli increase in PKD1 phosphorylation at Ser744/S748 without an increase in autophosphorylation at Ser916. This study exposes a mechanism that results in a discrepancy between PKD1 COOH-terminal autocatalytic activity and activity toward other substrates. We show that PKD1 constructs harboring COOH-terminal epitope tags display high levels of in vitro activation loop autocatalytic activity and activity toward ...