Lipid modulation of the activity of diacylglycerol kinase α- and ζ-isoforms: Activation by phosphatidylethanolamine and cholesterol

Maria Laura Fanani, Matthew K. Topham, James P. Walsh, Richard M. Epand

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Diacylglycerol kinase (DGK) isoforms α and ζ were extracted from transfected cells that overexpressed these enzymes. We determined the lipid dependence of the binding of these isoforms to liposomes. The modulation by lipid of the rate of phosphorylation of diacylglycerol by these enzymes was also measured. Incorporation of phosphatidylethanolamine into the liposomes resulted in an increased partitioning of both isoforms of DGK to the membrane as well as an increased catalytic rate. We demonstrate that the increased catalytic rate is a consequence of both increased portioning of the enzyme to the membrane and increased catalytic activity of the membrane-bound form. DGKα, a calcium-dependent isoform, can be activated in a calcium-independent fashion in the presence of phosphatidylethanolamine. Similar effects are observed with cholesterol. In contrast, sphingomyelin inhibits the activity of both isofonns of DGK. Our results demonstrate that the translocation to membranes and activity of DGKα and DGKζ are modulated by the composition and properties of the membrane. The enzymes are activated by the presence of lipids that promote the formation of inverted phases. However, the promotion of negative curvature is not the sole factor contributing to the lipid effects on enzyme binding and activity. A truncated form of DGKα lacking both the E-F hand and the recoverin homology domain is constitutively active and is not further activated by any of the lipids tested or by calcium. However, a truncated form lacking only the recoverin homology domain is partially activated by either calcium or certain lipids.

Original languageEnglish
Pages (from-to)14767-14777
Number of pages11
JournalBiochemistry
Volume43
Issue number46
DOIs
StatePublished - Nov 23 2004

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Diacylglycerol Kinase
Protein Isoforms
Chemical activation
Cholesterol
Modulation
Lipids
Membranes
Recoverin
Calcium
Enzymes
Liposomes
Phosphorylation
Sphingomyelins
Diglycerides
phosphatidylethanolamine
Catalyst activity
Hand
Chemical analysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lipid modulation of the activity of diacylglycerol kinase α- and ζ-isoforms : Activation by phosphatidylethanolamine and cholesterol. / Fanani, Maria Laura; Topham, Matthew K.; Walsh, James P.; Epand, Richard M.

In: Biochemistry, Vol. 43, No. 46, 23.11.2004, p. 14767-14777.

Research output: Contribution to journalArticle

Fanani, Maria Laura ; Topham, Matthew K. ; Walsh, James P. ; Epand, Richard M. / Lipid modulation of the activity of diacylglycerol kinase α- and ζ-isoforms : Activation by phosphatidylethanolamine and cholesterol. In: Biochemistry. 2004 ; Vol. 43, No. 46. pp. 14767-14777.
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