title: Dimethylarginine dimethylaminohydrolase activity modulates ADMA levels, VEGF expression, and cell phenotype.
abstract: Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase and is metabolised by dimethylarginine dimethylaminohydrolase (DDAH). Elevated levels of circulating ADMA correlate with various cardiovascular pathologies less is known about the cellular effects of altered DDAH activity. We modified DDAH activity in cells and measured the changes in ADMA levels, morphological phenotypes on Matrigel, and expression of vascular endothelial growth factor (VEGF). DDAH over-expressing ECV304 cells secreted less ADMA and when grown on Matrigel had enhanced tube formation compared to untransfected cells. VEGF mRNA levels were 2.1-fold higher in both ECV304 and murine endothelial cells (sEnd.1) over-expressing DDAH. In addition the DDAH inhibitor, S-2-amino-4(3-methylguanidino)butanoic acid (4124W 1mM), markedly reduced human umbilical vein endothelial cell tube formation in vitro. We have found that upregulating DDAH activity lowers ADMA levels, increases the levels of VEGF mRNA in endothelial cells, and enhances tube formation in an in vitro model, whilst blockade of DDAH reduces tube formation.
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, British Heart Foundation Laboratories, Department of Medicine, University College London, 5, University Street, London, UK. c.l.smith@ucl.ac.uk
authors.forename: Caroline L
authors.initials: CL
authors.lastname: Smith
authors.forename: Graeme M
authors.initials: GM
authors.lastname: Birdsey
authors.forename: Shelagh
authors.initials: S
authors.lastname: Anthony
authors.forename: Francesca I
authors.initials: FI
authors.lastname: Arrigoni
authors.forename: James M
authors.initials: JM
authors.lastname: Leiper
authors.forename: Patrick
authors.initials: P
authors.lastname: Vallance
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism*; D001120:Arginine / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000737:chemistry; D015152:Blotting, Northern; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D016228:Endothelial Growth Factors / Q000096:biosynthesis* / Q000737:chemistry; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D015151:Immunoblotting; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000737:chemistry; D007797:Laminin / Q000494:pharmacology; D008222:Lymphokines / Q000096:biosynthesis* / Q000737:chemistry; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D004338:Drug Combinations; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D008222:Lymphokines; D011509:Proteoglycans; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; C059485:matrigel; C018524:N,N-dimethylarginine; D003094:Collagen; D001120:Arginine; D000581:Amidohydrolases; C059999:dimethylargininase