The degradation of platelet-activating factor and related lipids: susceptibility to phospholipases C and D.
1-O-Octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3) is an ether-linked lipid that exhibits selective cytotoxicity toward several types of tumor cells and is relatively inactive toward normal cells under the same conditions of treatment. The mechanis of this selective cytotoxicity is unknown. We conducted studies to determine whether this compound is metabolized by phospholipases C and D and, if so, whether sensitive and resistant cells differ in their ability to degrade ET-18-OCH3 by these enzymes. We have examined the metabolism of the L-isomer of ET-18-OCH3, 1-O-octadecyl-2-O-methyl-sn-glycero-3-phosphocholine (L-ET-18-OCH3), by lysophospholipase D of rat liver microsomes and by a phospholipase D from the marine bacterium Vibrio damsela. The metabolism of L-ET-18-OCH3 was also examined in cell culture using Madin-Darby canine kidney cells, human promyelocytic leukemia cells and human myelocytic leukemia cells. In these studies, L-ET-18-OCH3 and related 1-O-alkyl-linked phosphocholine analogs radiolabeled with 3H in the 1-O-alkyl chain were used. L-ET-18-OCH3 was not hydrolyzed by lysophospholipase D from rat liver microsomes under conditions where cleavage of 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine was observed. However, phospholipase D from the marine bacterium V. damsela readily hydrolyzed L-ET-18-OCH3 to 1-O-[3H]octadecyl-2-O-methyl-sn-glycero-3-phosphate, demonstrating that L-ET-18-OCH3 can be degraded by a phospholipase D. Platelet-activating factor (PAF) and lyso-PAF were also substrates for the bacterial phospholipase D.(ABSTRACT TRUNCATED AT 250 WORDS)
Wilcox
R W
RW
Department of Biochemistry, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27103.
Wykle
R L
RL
Schmitt
J D
JD
Daniel
L W
LW
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism; D006801:Humans; D010739:Phospholipase D / Q000378:metabolism*; D010740:Phospholipases / Q000378:metabolism*; D010742:Phospholipid Ethers / Q000378:metabolism* / Q000493:pharmacokinetics; D010972:Platelet Activating Factor / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism*
D000970:Antineoplastic Agents; D010742:Phospholipid Ethers; D010972:Platelet Activating Factor; C026659:edelfosine; D010740:Phospholipases; D010738:Type C Phospholipases; D010739:Phospholipase D