title: Human umbilical vein endothelial cells generate leukotriene C4 via microsomal glutathione S-transferase type 2 and express the CysLT(1) receptor.
abstract: Certain immunocompetent myeloid cells, such as eosinophils, basophils and mast cells, have a large capacity to synthesize the potent proinflammatory and spasmogenic mediator leukotriene (LT) C4 via a specific microsomal glutathione S-transferase (MGST) termed LTC4 synthase (LTC4S). Here, we report that MGST2, a distant homologue of LTC4S, is abundantly expressed in Human umbilical vein endothelial cells (HUVEC) and converts LTA4 into a single product, LTC4. Thus, using Northern blot, RT-PCR, Western blot, and enzyme activity assays, we show that MGST2 is the main, if not the only, enzyme that converts LTA4 into LTC4 in membrane preparations of HUVEC. In fact, we failed to detect any expression of LTC4S, MGST1 or MGST3 in these cells, indicating that MGST2 is a critical enzyme for transcellular LTC4 biosynthesis in the vascular wall. Unlike LTC4S, MGST2 prefers the naturally occurring free acid of LTA4 over the methyl ester as substrate and is also susceptible to product inhibition with an IC50 of about 1 microM for LTC4. Moreover, HUVEC were found to express the CysLT1 receptor in line with a paracrine and autocrine role for cysteinyl-leukotrienes in endothelial cell function.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Division of Chemistry II, Karolinska Institutet, Stockholm, Sweden.
authors.forename: M
authors.initials: M
authors.lastname: Sjöström
authors.forename: P J
authors.initials: PJ
authors.lastname: Jakobsson
authors.forename: M
authors.initials: M
authors.lastname: Heimburger
authors.forename: J
authors.initials: J
authors.lastname: Palmblad
authors.forename: J Z
authors.initials: JZ
authors.lastname: Haeggström
mesh_terms: D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D017572:Leukotriene A4 / Q000378:metabolism; D017997:Leukotriene C4 / Q000096:biosynthesis* / Q000494:pharmacology; D008565:Membrane Proteins; D008861:Microsomes / Q000201:enzymology; D008954:Models, Biological; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018077:Receptors, Leukotriene / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D014471:Umbilical Veins / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D017572:Leukotriene A4; D008565:Membrane Proteins; D012333:RNA, Messenger; D018077:Receptors, Leukotriene; D011994:Recombinant Proteins; D017997:Leukotriene C4; C412375:cysteinyl leukotriene receptor 2; D005982:Glutathione Transferase; C041116:leukotriene-C4 synthase; C083169:leukotriene D4 receptor