Golgi-associated protein kinase C-ϵ is delivered to phagocytic cups: Role of phosphatidylinositol 4-phosphate

Cheryl M. Hanes, Anna E. D'Amico, Takehiko Ueyama, Alexander C. Wong, Xuexin Zhang, W. Frederick Hynes, Margarida M. Barroso, Nathaniel C. Cady, Mohamed Trebak, Naoaki Saito, Michelle R. Lennartz

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Protein kinase C-ϵ (PKC-ϵ) at phagocytic cups mediates the membrane fusion necessary for efficient IgG-mediated phagocytosis. The C1B and pseudosubstrate (ϵPS) domains are necessary and sufficient for this concentration. C1B binds diacylglycerol; the docking partner for ϵPS is unknown. Liposome assays revealed that the ϵPS binds phosphatidylinositol 4-phosphate (PI4P) and PI(3,5)P2. Wortmannin, but not LY294002, inhibits PKC-ϵ concentration at cups and significantly reduces the rate of phagocytosis. As Wortmannin inhibits PI4 kinase, we hypothesized that PI4P mediates the PKC-ϵ concentration at cups and the rate of phagocytosis. PKC-ϵ colocalizes with the trans-Golgi network (TGN) PI4P reporter, P4M, suggesting it is tethered at the TGN. Real-time imaging of GFP-PKC-ϵ-expressing macrophages revealed a loss of Golgi-associated PKC-ϵ during phagocytosis, consistent with a Golgi-to-phagosome translocation. Treatment with PIK93, a PI4 kinase inhibitor, reduces PKC-ϵ at both the TGN and the cup, decreases phagocytosis, and prevents the increase in capacitance that accompanies membrane fusion. Finally, expression of the Golgi-directed PI4P phosphatase, hSac1-K2A, recapitulates the PIK93 phenotype, confirming that Golgiassociated PI4P is critical for efficient phagocytosis. Together these data are consistent with a model in which PKC-ϵ is tethered to the TGN via an ϵPS-PI4P interaction. The TGN-associated pool of PKC-ϵ concentrates at the phagocytic cup where it mediates the membrane fusion necessary for phagocytosis. The novelty of these data lies in the demonstration that ϵPS binds PI4P and PI(3,5)P2 and that PI4P is necessary for PKC-ϵ localization at the TGN, its translocation to the phagocytic cup, and the membrane fusion required for efficient Fc [γ] receptor-mediated phagocytosis.

Original languageEnglish (US)
Pages (from-to)271-277
Number of pages7
JournalJournal of Immunology
Issue number1
StatePublished - Jul 1 2017

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology


Dive into the research topics of 'Golgi-associated protein kinase C-ϵ is delivered to phagocytic cups: Role of phosphatidylinositol 4-phosphate'. Together they form a unique fingerprint.

Cite this