Catabolism of platelet-activating factor by human colonic mucosa. Calcium dependence of the catabolizing enzymes.
The catabolism of platelet-activating factor (PAF) and lyso PAF by a supernatant fraction of human colon mucosa homogenates has been studied in vitro. PAF is initially catabolized to lyso PAF by mucosal enzymes via removal of its acetyl group. Incubates in Ca(2+)-free Tris with EDTA showed that the acetyl hydrolase was Ca2+ independent. Addition of the hydrolase inhibitor, phenyl methyl sulphonyl fluoride, significantly reduced the catabolism of PAF. Lyso PAF was further catabolized in at least two ways. An acyl group was incorporated into the sn-2 position of lyso PAF to give 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine (alkyl acyl GPC); this step was Ca2+ independent as shown by omitting Ca2+ and adding EDTA to the incubate. Formation of alkyl acyl GPC was confirmed by HPLC. Alternatively, choline was removed from the head group of lyso PAF by a calcium-dependent lyso phospholipase D. Under the experimental conditions utilized a neutral lipid product was formed but significant amounts of the intermediate lysophosphatidic acid could not be detected. A substance with a chromatographic mobility of Rf = 0.8 on TLC plates having an intact phosphorylcholine head group was also formed but has not yet been identified. It is concluded that the human colon mucosa contains enzymes that actively catabolize pro-inflammatory PAF and lyso PAF.
Appleyard
C B
CB
Clinical Pharmacology Group, Medical Faculty, University of Southampton, U.K.
Hillier
K
K
D002118:Calcium / Q000494:pharmacology*; D003106:Colon / Q000201:enzymology*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000378:metabolism*; D013997:Time Factors
C069030:1-acyl-2-acetyl-sn-glycero-3-phosphocholine; C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; D010664:Phenylmethylsulfonyl Fluoride; D002118:Calcium