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The Journal of Lipid Research, Vol. 39, 569-582, March 1998
Copyright © 1998 by Lipid Research, Inc.

Article

Identification and characterization of several forms of phospholipase A₂ in mouse epidermal keratinocytes

Correspondence to: Susan M. Fischer.

The tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA) stimulated the release of arachidonic acid (AA) from mouse keratinocytes. A distinct difference was observed between the fatty acid release profile elicited by TPA and other stimuli. These findings led to the investigation of keratinocyte phospholipase A₂ (PLA₂), which catalyzes the release of sn-2 fatty acids from membrane phospholipids and regulates the production of eicosanoids. We characterized and identified several forms of PLA₂ in mouse keratinocytes, a cytosolic or cPLA₂ and two secretory or sPLA₂s in the membrane. The PLA₂ in keratinocyte cytosol is sensitive to heating and acid treatment, while resistant to reducing reagent. The PLA₂ in keratinocyte membrane is resistant to heating and acid treatment, while sensitive to reducing reagent. These characteristics suggested the presence of a cPLA ₂ and at least one type of sPLA₂. Inhibitor data further confirmed the identities of these PLA₂s. The cPLA₂ was activated by TPA, and appeared to be responsible for the majority of the specific release of AA observed in mouse keratinocytes treated with TPA. The calcium ionophore A23187, and 4-TPA did not elicit the selectivity towards AA observed with TPA. The release of linoleic acid (LA) and oleic acid (OA) from A23187- and 4-TPA-treated keratinocytes suggests activation of sPLA₂. These activities may be due to the existence of both type I and type II sPLA₂, as both were identified by polymerase chain reactions.

In conclusion, keratinocytes express several forms of phospholipase A ₂ that differ in their substrate specificities and mechanisms of activation, resulting in distinct agonist-specific fatty acid release profiles.—Li-Stiles, B., H-H. Lo, and S. M. Fischer. Identification and characterization of several forms of phospholipase A ₂ in mouse epidermal keratinocytes. J. Lipid Res. 1998. 39: 569–582.

Supplementary key words: arachidonic acid, cPLA₂, keratinocytes, PLA₂, sPLA₂, TPA

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