Palmitoylation controls the catalytic activity and subcellular distribution of phosphatidylinositol 4-kinase IIα

Barbara Barylko, Yuntao S. Mao, Pawel Wlodarski, Gwanghyun Jung, Derk D. Binns, Hui Qiao Sun, Helen L. Yin, Joseph P. Albanesi

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Phosphatidylinositol 4-kinases play essential roles in cell signaling and membrane trafficking. They are divided into type II and III families, which have distinct structural and enzymatic properties and are essentially unrelated in sequence. Mammalian cells express two type II isoforms, phosphatidylinositol 4-kinase IIα (PI4KIIα) and IIβ (PI4KIIβ). Nearly all of PI4KIIα, and about half of PI4KIIβ, associates integrally with membranes, requiring detergent for solubilization. This tight membrane association is because of palmitoylation of a cysteine-rich motif, CCPCC, located within the catalytic domains of both type II isoforms. Deletion of this motif from PI4KIIα converts the kinase from an integral to a tightly bound peripheral membrane protein and abrogates its catalytic activity (Barylko, B., Gerber, S. H., Binns, D. D., Grichine, N., Khvotchev, M., Sudhof, T. C., and Albanesi, J. P. (2001) J. Biol. Chem. 276, 7705-7708). Here we identify the first two cysteines in the CCPCC motif as the principal sites of palmitoylation under basal conditions, and we demonstrate the importance of the central proline for enzymatic activity, although not for membrane binding. We further show that palmitoylation is critical for targeting PI4KIIα to the trans-Golgi network and for enhancement of its association with low buoyant density membrane fractions, commonly termed lipid rafts. Replacement of the four cysteines in CCPCC with a hydrophobic residue, phenylalanine, substantially restores catalytic activity of PI4KIIα in vitro and in cells without restoring integral membrane binding. Although this FFPFF mutant displays a perinuclear distribution, it does not strongly co-localize with wild-type PI4KIIα and associates more weakly with lipid rafts.

Original languageEnglish (US)
Pages (from-to)9994-10003
Number of pages10
JournalJournal of Biological Chemistry
Volume284
Issue number15
DOIs
StatePublished - Apr 10 2009

Fingerprint

Lipoylation
1-Phosphatidylinositol 4-Kinase
Catalyst activity
Membranes
Cysteine
Protein Isoforms
Cell signaling
Lipids
trans-Golgi Network
Cell membranes
Phenylalanine
Proline
Detergents
Membrane Proteins
Phosphotransferases
Catalytic Domain
Cells
Cell Membrane

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Palmitoylation controls the catalytic activity and subcellular distribution of phosphatidylinositol 4-kinase IIα. / Barylko, Barbara; Mao, Yuntao S.; Wlodarski, Pawel; Jung, Gwanghyun; Binns, Derk D.; Sun, Hui Qiao; Yin, Helen L.; Albanesi, Joseph P.

In: Journal of Biological Chemistry, Vol. 284, No. 15, 10.04.2009, p. 9994-10003.

Research output: Contribution to journalArticle

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AU - Barylko, Barbara

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AU - Binns, Derk D.

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