title: N-domain angiotensin-converting enzyme isoform expression in tissues of Wistar and spontaneously hypertensive rats.
abstract: BACKGROUND
  Angiotensin I-converting enzyme (ACE) is a protein containing two active sites, called N- and C-domains, according to their position in the protein.
  
  
  OBJECTIVE
  The aim of the present study was to verify whether the expression of the N-domain ACEs detected in the urine of Wistar and spontaneously hypertensive (SHR) rats was restricted to the kidney.
  
  
  METHODS
  Adrenal, aorta, heart, liver, lung, kidney and testicle tissue from Wistar rats and spontaneously hypertensive rats were homogenized in assay buffer and analyzed by gel filtration, Western blotting and radio-immunoassay.
  
  
  RESULTS
  Two peaks (at 136 and 69 kDa) with ACE activity upon ZPhe-His-Leu were separated by gel filtration from homogenate tissues of Wistar rats, in contrast with the tissue from hypertensive rats, which showed ACE forms of 96 and 69 kDa. The bands detected by Western blotting for all studied tissue from Wistar and spontaneously hypertensive rats showed a correspondence with the two peaks containing ACE activity detected in the polyacrylamide gel slices. Angiotensin II levels were increased in hypertensive rat tissue when compared with Wistar rat tissues. In addition, captopril 3 micromol/l inhibited the enzymic activity, where the Km was in the order of mmol/l and micromol/l using hippuryl-His-Leu and Abz-Ser-Asp-Lys(Dnp)Pro-OH as substrates, respectively. All tissues from Wistar rats presented ACE with 136 kDa, similar to somatic ACE, and N-domain ACE with 69 kDa. In the same tissue of spontaneously hypertensive rats, 96 and 69 kDa N-domain ACEs were detected.
  
  
  CONCLUSIONS
  Our results demonstrated that N-domain ACEs were not exclusively produced in the kidney and excreted in the urine; they were expressed in all tissue studied, suggesting that these enzymes could influence local angiotensin II production, contributing to organ-specific regulation.
authors.affiliation: Nephrology Division, Department of Medicine, Federal University of São Paulo, Brazil.
authors.forename: Fernanda A
authors.initials: FA
authors.lastname: Ronchi
authors.forename: Maria C C
authors.initials: MC
authors.lastname: Andrade
authors.forename: Adriana K
authors.initials: AK
authors.lastname: Carmona
authors.forename: José E
authors.initials: JE
authors.lastname: Krieger
authors.forename: Dulce E
authors.initials: DE
authors.lastname: Casarini
mesh_terms: D000311:Adrenal Glands / Q000201:enzymology; D000804:Angiotensin II / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000201:enzymology; D001665:Binding Sites; D015153:Blotting, Western; D002850:Chromatography, Gel; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D009206:Myocardium / Q000201:enzymology; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D011918:Rats, Inbred SHR; D017208:Rats, Wistar; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology
chemical_list: D007527:Isoenzymes; D000804:Angiotensin II; D007703:Peptidyl-Dipeptidase A