title: Characterization of chicken lung angiotensin I-converting enzyme.
abstract: Angiotensin I(AI)-converting enzyme (ACE) (EC 3.4.15.1) was solubilized from the membrane fraction of chicken lung using trypsin and nonidet P40 extraction, and then purified to homogeneity by captopril affinity chromatography. Comparison of trypsin-extracted and detergent-solubilized membrane-bound converting enzyme by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and isoelectric focusing indicated that the membrane-binding sequence contributed to a large extent to the size and charge of the enzyme. Both forms of the enzyme were glycoproteins but they differed in the glucidic content; 4.5% by weight of the enzyme in the trypsin-extracted ACE and 15% by weight of the enzyme in the detergent-solubilized ACE. In both cases hexoses were the most abundant residues. Both forms of the enzyme were found to contain 1 g-atom zinc/mol enzyme. The purified enzymes did not only split Hip-His-Leu but also AI and bradykinin. The Michaelis constant (Km) and maximum velocity (Vmax) values of the trypsin-extracted ACE for Hip-His-Leu were 52 x 10(-5) mol/l and 15.36 nmol/min respectively, and for AI they were 7.8 x 10(-5) mol/l and 0.45 nmol/min respectively. The Km and Vmax values of the detergent-solubilized ACE for Hip-His-Leu were 32 x 10(-5) mol/l and 11.75 nmol/min respectively, and for AI they were 6.5 x 10(-5) mol/l and 0.97 nmol/min.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Departamento de Bioquímica, Facultad de Ciencias, Universidad de Valladolid, Spain.
authors.forename: M J
authors.initials: MJ
authors.lastname: Polanco
authors.forename: J L
authors.initials: JL
authors.lastname: Miguel
authors.forename: M T
authors.initials: MT
authors.lastname: Agapito
authors.forename: J M
authors.initials: JM
authors.lastname: Recio
mesh_terms: D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002645:Chickens / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008168:Lung / Q000737:chemistry*; D007703:Peptidyl-Dipeptidase A / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D015032:Zinc / Q000032:analysis
chemical_list: D002241:Carbohydrates; D007703:Peptidyl-Dipeptidase A; D015032:Zinc