title: Immunolocalisation and activity of DDAH I and II in the heart and modification post-myocardial infarction.
abstract: Asymmetric dimethylarginine (ADMA) and N(G) monomethyl-L-arginine (L-NMMA) are endogenous inhibitors of nitric oxide synthases (NOS) and their local concentration is determined by the activity of dimethylarginine dimethylaminohydrolases (DDAHs). The current study in male Wistar rats was designed to immunolocalise DDAH I and II in relation to NOS and to investigate changes in distribution, activity and ADMA content in the acute period following myocardial infarction (MI) resulting from coronary artery ligation. Seven days after the coronary artery ligation, L-Arg and methylated arginine content, as well as DDAH activity were determined in homogenates of left ventricular (LV) infarct and border. The distribution of immunoreactive DDAH I, DDAH II, eNOS and iNOS were determined in sections of LV. In healthy hearts, DDAH I was absent, however, DDAH II was localized to endothelium and endocardium with a similar distribution to that of eNOS. Following MI, LV DDAH activity was increased (to 210+/-19% of control, P<0.05). Both DDAH I and DDAH II proteins were detected in peri-infarct cardiomyocytes, while DDAH II immunoreactivity was additionally localized to infiltrating inflammatory cells and blood vessels in the healing infarct. Both plasma and LV concentrations of the DDAH substrate, ADMA, were increased post-MI, although the ratio of Arg:ADMA was retained in the LV post-MI relative to sham operated controls. In conclusion, DDAH II has a distribution similar to eNOS in healthy myocardium. The increased levels and activity of DDAH I and DDAH II enzymes following myocardial infarction suggest a potential role for them in local protection of NOS enzymes from inhibition by methylated arginines during infarct healing.
authors.affiliation: Centre for Cardiovascular Science, Queens Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh EH16 4TJ, UK. gillian.gray@ed.ac.uk
authors.forename: Gillian A
authors.initials: GA
authors.lastname: Gray
authors.forename: Mark
authors.initials: M
authors.lastname: Patrizio
authors.forename: Lorcan
authors.initials: L
authors.lastname: Sherry
authors.forename: Alyson A
authors.initials: AA
authors.lastname: Miller
authors.forename: Mohammed
authors.initials: M
authors.lastname: Malaki
authors.forename: Alison F
authors.initials: AF
authors.lastname: Wallace
authors.forename: James M
authors.initials: JM
authors.lastname: Leiper
authors.forename: Patrick
authors.initials: P
authors.lastname: Vallance
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000201:enzymology; D001120:Arginine / Q000031:analogs & derivatives / Q000097:blood / Q000378:metabolism; D004195:Disease Models, Animal; D004699:Endocardium / Q000201:enzymology; D006352:Heart Ventricles / Q000201:enzymology / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D008297:Male; D009203:Myocardial Infarction / Q000097:blood / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D009206:Myocardium / Q000201:enzymology* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000201:enzymology; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
chemical_list: C487735:dimethylarginine; C024917:symmetric dimethylarginine; D001120:Arginine; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496318:Nos2 protein, rat; C496316:Nos3 protein, rat; C541817:PRMT1 protein, rat; D011484:Protein-Arginine N-Methyltransferases; D000581:Amidohydrolases; C059999:dimethylargininase