Mechanisms that account for the selective release of arachidonic acid from intact cells by secretory phospholipase A2.
The current study examined mechanisms that account for the selective release of arachidonic acid (AA) from cells by secretory phospholipase A2 (sPLA2). Initial studies demonstrated that low concentrations of group I and group III PLA2 isotypes and an sPLA2-enriched extract from bone marrow-derived mast cells (BMMC) selectively released AA from mast cells. Much higher concentrations of group II PLA2 were required to release comparable quantities of AA. Group I PLA2 also selectively released AA from another mast cell line (CFTL-15) and a monocytic cell line (THP-1). In contrast, high concentrations of group I PLA2 were required to release fatty acids from a promyelocytic cell line (HL-60) and this release was not selective for AA. Binding studies revealed that cell types (BMMC, CFTL-15 and THP-1) which selectively released AA also had the capacity to specifically bind group I PLA2. However, group II PLA2, which did not selectively release AA from cells, also did not specifically bind to these same cell types. Additional studies revealed that sPLA2 binding to the mast cell receptor was attenuated after stimulation with antigen or ionophore A23187. Reverse transcriptase-polymerase chain reaction analyses indicated the presence of mRNA for the sPLA2 receptor in BMMC, CFTL-15 and THP-1 and the absence of this mRNA in HL-60. Final studies demonstrated that p-aminophenyl-alpha-D-mannopyranoside BSA, a known ligand of the sPLA2 receptor, also selectively released AA from mast cells but not from HL-60 cells. These experiments indicated that receptor occupancy alone (without PLA2 activity) is sufficient to induce the release of AA from mast cells. Together, these data reveal that specific isotypes of sPLA2 have the capacity to selectively release AA from certain cells by their capacity to bind to sPLA2 receptors on the cell surface.
Fonteh
A N
AN
Department of Internal Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA. afonteh@wfubmc.edu
Samet
J M
JM
Surette
M
M
Reed
W
W
Chilton
F H
FH
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000201:enzymology / Q000378:metabolism*; D002460:Cell Line; D005227:Fatty Acids / Q000032:analysis; D054501:Group II Phospholipases A2; D007457:Iodine Radioisotopes; D007527:Isoenzymes / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D008808:Mice, Inbred CBA; D010741:Phospholipases A / Q000378:metabolism / Q000494:pharmacology*; D054467:Phospholipases A2; D011956:Receptors, Cell Surface / Q000378:metabolism
D005227:Fatty Acids; D007457:Iodine Radioisotopes; D007527:Isoenzymes; D011956:Receptors, Cell Surface; D016718:Arachidonic Acid; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2