Naegleria fowleri amoebae express a membrane-associated calcium-independent phospholipase A(2).
Naegleria fowleri, a free-living amoeba, is the causative agent of primary amoebic meningoencephalitis. Previous reports have demonstrated that N. fowleri expresses one or more forms of phospholipase A(2) (PLA(2)) and that a secreted form of this enzyme is involved in pathogenesis. However, the molecular nature of these phospholipases remains largely unknown. This study was initiated to determine whether N. fowleri expresses analogs of the well-characterized PLA(2)s that are expressed by mammalian macrophages. Amoeba cell homogenates contain a PLA(2) activity that hydrolyzes the substrate that is preferred by the 85 kDa calcium-dependent cytosolic PLA(2), cPLA(2). However, unlike the cPLA(2) enzyme in macrophages, this activity is largely calcium-independent, is constitutively associated with membranes and shows only a modest preference for phospholipids that contain arachidonate. The amoeba PLA(2) activity is sensitive to inhibitors that block the activities of cPLA(2)-alpha and the 80 kDa calcium-independent PLA(2), iPLA(2), that are expressed by mammalian cells. One of these compounds, methylarachidonyl fluorophosphonate, partially inhibits the constitutive release of [(3)H]arachidonic acid from pre-labeled amoebae. Together, these data suggest that N. fowleri expresses a constitutively active calcium-independent PLA(2) that may play a role in the basal phospholipid metabolism of these cells.
Department of Microbiology and Immunology, Virginia Commonwealth University, Box 980678, Richmond, VA 23298-0678, USA. sbarbour@hsc.vcu.edu
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Marciano-Cabral
D000818:Animals; D016718:Arachidonic Acid / Q000032:analysis / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D054522:Group VI Phospholipases A2; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D016848:Naegleria fowleri / Q000201:enzymology / Q000378:metabolism*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D014316:Tritium
D010743:Phospholipids; D014316:Tritium; D016718:Arachidonic Acid; D010741:Phospholipases A; D054522:Group VI Phospholipases A2; C516476:Pla2g6 protein, mouse