Movement of monoglyceride derived from hydrolysis of fluorescence-labeled lyso platelet-activating factor by lysophospholipase C through plasma membranes of porcine kidney epithelial cell line LLC-PK1.
To investigate the mechanisms of the release of lyso platelet-activating factor (PAF), an alkyl ether-linked lysophosphatidylcholine, from the kidney epithelial cell line LLC-PK1, the cell monolayer was incubated with a fluorescence-labeled lysoPAF analog, Bodipy-lysoPAF, on either the basolateral or apical side. The fluorescent lipids in the culture media mixed with or without bovine serum albumin at a final concentration of 2% were analyzed by thin layer chromatography. In both cases, two major bands, assignable to Bodipy-lysoPAF and Bodipy-monoglyceride (MG), were detected in the culture medium to which Bodipy-lysoPAF had been added, whereas the culture medium at the opposite side exhibited only the major band of Bodipy-MG. Our results suggest that lysoPAF was degraded by high ecto-lysophospholipase C activity. The possible physiological significance of this metabolic pathway is discussed.
Tsutsumi
Toshihiko
T
Faculty of Pharmaceutical Sciences, Kyushu University of Health and Welfare, 1714-1 Yoshinocho, Nobeoka 882-8508, Japan.
Morishige
Junichi
J
Fukuzawa
Kenji
K
Tokumura
Akira
A
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D001896:Boron Compounds / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology*; D002855:Chromatography, Thin Layer; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005997:Glycerylphosphorylcholine / Q000494:pharmacology; D006868:Hydrolysis / Q000187:drug effects; D007668:Kidney / Q000166:cytology* / Q000187:drug effects; D008245:Lysophospholipase / Q000378:metabolism*; D050178:Monoglycerides / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism; D013552:Swine
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D050178:Monoglycerides; C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; D005997:Glycerylphosphorylcholine; D008245:Lysophospholipase