title: The primary determinant of rabbit myocardial ethanolamine phosphotransferase substrate selectivity is the covalent nature of the sn-1 aliphatic group of diradyl glycerol acceptors.
abstract: Plasmenylethanolamines represent the major endogenous phospholipid storage depot of arachidonic acid in many mammalian cells. To elucidate the biochemical mechanisms contributing to the high plasmalogen content and arachidonic acid enrichment present in myocardial ethanolamine glycerophospholipids, the substrate specificity of rabbit myocardial ethanolamine phosphotransferase (EPT) was quantified utilizing multiple molecular species of each subclass of diradyl glycerol substrate. Myocardial EPT demonstrated over a 16-fold selectivity for 1-O-alk-1'-enyl-2-acyl-sn-glycerol (AAG) compared to 1,2-diacyl-sn-glycerol (DAG) substrate utilizing individual molecular species of each subclass dispersed in Tween 20. The selective utilization of AAG by EPT was substantiated utilizing two independent assay systems which employed either the presentation of substrate to enzyme as a substitutional impurity in Triton X-100 mixed micelles or the obligatory utilization of endogenously generated diradyl glycerol substrates. Although rabbit myocardial microsomes contained over a 20-fold molar excess of endogenous DAG to AAG mass, incubation of rabbit myocardial microsomes with CDP-ethanolamine resulted in the highly selective synthesis of plasmenylethanolamines which were predominantly comprised of molecular species containing arachidonic acid at the sn-2 position (greater than 75%). Endogenous AAG molecular species in rabbit myocardial microsomes were similarly enriched in arachidonic acid, and the distribution of AAG molecular species closely paralleled the distribution of plasmenylethanolamine (but not plasmenylcholine) molecular species. Thus, the subclass and molecular species distribution of the ethanolamine glycerophospholipids synthesized by rabbit myocardial EPT reflects independent contributions from the subclass selectivity of EPT for AAG substrate in conjunction with the enrichment of arachidonic acid in microsomal AAG molecular species.
authors.affiliation: Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: D A
authors.initials: DA
authors.lastname: Ford
authors.forename: K B
authors.initials: KB
authors.lastname: Rosenbloom
authors.forename: R W
authors.initials: RW
authors.lastname: Gross
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D004982:Ethanolaminephosphotransferase / Q000378:metabolism*; D005990:Glycerol / Q000378:metabolism*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology*; D010955:Plasmalogens / Q000096:biosynthesis; D011817:Rabbits; D013379:Substrate Specificity
chemical_list: D010955:Plasmalogens; C020791:phosphatidal ethanolamines; D016718:Arachidonic Acid; D004982:Ethanolaminephosphotransferase; D005990:Glycerol