PKN Regulates Phospholipase D1 through Direct Interaction

• Published in: The Journal of biological chemistry 2001-05, Vol.276 (21), p.18096-18101
• Main Authors: Oishi, Kumiko, Takahashi, Mikiko, Mukai, Hideyuki, Banno, Yoshiko, Nakashima, Shigeru, Kanaho, Yasunori, Nozawa, Yoshinori, Ono, Yoshitaka
• Format: Article
• Language: English
• Subjects: Animals; COS Cells; Enzyme Activation; HeLa Cells; Humans; Phospholipase D - genetics; Phospholipase D - metabolism; Protein Binding; Protein Kinase C; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - metabolism; Protein-Tyrosine Kinases - genetics; Protein-Tyrosine Kinases - metabolism; Signal Transduction
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M010646200

The association of phospholipase (PLD)-1 with protein kinase C-related protein kinases, PKNα and PKNβ, was analyzed. PLD1 interacted with PKNα and PKNβ in COS-7 cells transiently transfected with PLD1 and PKNα or PKNβ expression constructs. The interactions between endogenous PLD1 and PKNα or PKNβ were confirmed by co-immunoprecipitation from mammalian cells.In vitro binding studies using the deletion mutants of PLD1 indicated that PKNα directly bound to residues 228–598 of PLD1 and that PKNβ interacted with residues 1–228 and 228–598 of PLD1. PKNα stimulated the activity of PLD1 in the presence of phosphatidylinositol 4,5-bisphosphate in vitro, whereas PKNβ had a modest effect on the stimulation of PLD1 activity. The stimulation of PLD1 activity by PKNα was slightly enhanced by the addition of arachidonic acid. These results suggest that the PKN family functions as a novel intracellular player of PLD1 signaling pathway.