Arachidonoyl-diacylglycerol kinase

Specific in vitro inhibition by polyphosphoinositides suggests a mechanism for regulation of phosphatidylinositol biosynthesis

James P. Walsh, Rosa Suen, John A. Glomset

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

We previously described the purification of a membrane-bound diacylglycerol kinase highly selective for sn-1-acyl-2-arachidonoyl diacylglycerols (Walsh, J. P., Suen, R., Lemaitre, R. N., and Glomset, J. A. (1994) J. Biol. Chem. 269, 21155-21164). This enzyme appears to be responsible for the rapid clearance of the arachidonaterich pool of diacylglycerols generated during stimulusinduced phosphoinositide turnover. We have now shown phosphatidylinositol 4,5-bisphosphate to be a potent and specific inhibitor of arachidonoyl-diacylglycerol kinase. Kinetic analyses indicated a Ki for phosphatidylinositol 4,5-bisphosphate of 0.04 mol %. Phosphatidic acid also was an inhibitor with a Ki of 0.7 mol %. Other phospholipids had only small effects at these concentrations. A series of multiply phosphorylated lipid analogs also inhibited the enzyme, indicating that the head group phosphomonoesters are the primary determinants of the polyphosphoinositide effect. However, these compounds were not as potent as phosphatidylinositol 4,5-bisphosphate, indicating some specificity for the polyphosphoinositide additional to its total charge. Five other diacylglycerol kinases were activated to varying degrees by phosphatidylinositol 4,5-bisphosphate and phosphatidic acid, suggesting that inhibition by acidic lipids may be specific for the arachidonoyl-DAG kinase isoform. Given the presumed role of arachidonoyl-diacylglycerol kinase in the phosphoinositide cycle, this inhibition may represent a mechanism for polyphosphoinositides to regulate their own synthesis.

Original languageEnglish
Pages (from-to)28647-28653
Number of pages7
JournalJournal of Biological Chemistry
Volume270
Issue number48
StatePublished - Dec 1 1995

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Diacylglycerol Kinase
Phosphatidylinositol Phosphates
Biosynthesis
Phosphatidylinositols
Phosphatidic Acids
Diglycerides
Lipids
Enzymes
Purification
In Vitro Techniques
Phospholipids
Protein Isoforms
Membranes
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Arachidonoyl-diacylglycerol kinase : Specific in vitro inhibition by polyphosphoinositides suggests a mechanism for regulation of phosphatidylinositol biosynthesis. / Walsh, James P.; Suen, Rosa; Glomset, John A.

In: Journal of Biological Chemistry, Vol. 270, No. 48, 01.12.1995, p. 28647-28653.

Research output: Contribution to journalArticle

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