title: Competitive enzymatic interactions determine the relative amounts of prostaglandins E2 and D2.
abstract: Prostaglandins (PGs) are a family of cellular messengers exerting diverse homeostatic and pathophysiologic effects. Recently, several studies reported significant increases of PGI(2) and PGF(2α) after the inhibition of microsomal PGE synthase-1 (mPGES-1) expression, which indicated that PGH(2) metabolism might be redistributed when the PGE(2) pathway is blocked. To address the determinants that govern the relative amounts of PGs, we developed an in vitro cell-free method, based on liquid chromatography-tandem mass spectrometry, to measure the exact amounts of these PGs formed in response to the addition of recombinant isomerases and their selective inhibitors. Our in vitro cell-free assay results were confirmed in cells using bone marrow-derived macrophage. Initially, we determined the in vitro stability of PGH(2) and noted that there was spontaneous nonenzymatic conversion to PGD(2) and PGE(2). mPGES-1 markedly increased the conversion to PGE(2) and decreased conversion to PGD(2). Reciprocally, the addition of hematopoietic or lipocalin PGD synthase resulted in a relative increase of PGD(2) and decrease of PGE(2). A detailed titration study showed that the ratio of PGE(2)/PGD(2) was closely correlated with the ratio of PGE synthase/PGD synthase. Our redistribution results also provide the foundation for understanding how PGH(2) metabolism is redistributed by the presence of distal isomerases or by blocking the major metabolic outlet, which could determine the relative benefits and risks resulting from interdiction in nonrated-limiting components of PG synthesis pathways.
authors.affiliation: Department of Medicinal Chemistry and Pharmacognosy, University of Illinois College of Pharmacy, 833 S. Wood Street, Chicago, IL 60612-7231, USA.
authors.forename: Rui
authors.initials: R
authors.lastname: Yu
authors.forename: Lei
authors.initials: L
authors.lastname: Xiao
authors.forename: Guiqing
authors.initials: G
authors.lastname: Zhao
authors.forename: John W
authors.initials: JW
authors.lastname: Christman
authors.forename: Richard B
authors.initials: RB
authors.lastname: van Breemen
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D051545:Cyclooxygenase 1 / Q000378:metabolism*; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D003986:Dibenzocycloheptenes / Q000494:pharmacology; D015232:Dinoprostone / Q000378:metabolism*; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism; D007535:Isomerases / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology; D054834:Lipocalins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010880:Piperidines / Q000494:pharmacology; D015230:Prostaglandin D2 / Q000378:metabolism*; D044262:Prostaglandin H2 / Q000378:metabolism*; D000071876:Prostaglandin-E Synthases; D012756:Sheep; D013876:Thiophenes / Q000494:pharmacology
chemical_list: C569292:4-(5H-dibenzo(a,d)cyclohepten-5-ylidene)-1-(4-(2H-tetrazol-5-yl)butyl)piperidine; C569291:4-(benzo(b)thiophen-2-yl)-3-bromo-5-hydroxydihydrofuran-2(3H)-one; C116322:4-benzhydryloxy-1-(3-(1H-tetrazol-5-yl-)-propyl)piperidine; D003986:Dibenzocycloheptenes; D004791:Enzyme Inhibitors; D054834:Lipocalins; D010880:Piperidines; D013876:Thiophenes; D044262:Prostaglandin H2; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C545516:HPGDS protein, mouse; C022466:prostaglandin R2 D-isomerase; C000605862:PTGES protein, human; D000071876:Prostaglandin-E Synthases; C000605865:Ptges protein, mouse; D015232:Dinoprostone; D015107:4-Butyrolactone; D015230:Prostaglandin D2