Purification of angiotensin I converting enzyme from pig lung using concanavalin-A sepharose chromatography.
Angiotensin I converting enzyme (ACE) plays a major role in blood pressure regulation, catalyzing the conversion of angiotensin I to the vasoconstrictor angiotensin II. In this report we describe a two-step affinity chromatography method for preparative purification of ACE from pig lung using Concanavalin-A Sepharose 4B and affinity chromatography on Lisinopril Sepharose 6B. The same purification scheme was used to obtain Cobalt-ACE, where zinc ion located at the active site is replaced by cobalt. Cobalt-ACE visible spectrum shows a characteristic broad peak from 500 to 600 nm. The shape and maximum absorptivity of this peak changes in presence of ACE inhibitors that bind at the catalytic site.
Andujar-Sánchez
M
M
Dpto. Química Física, Bioquímica y Qui;mica Inorgánica, Universidad de Almería, Carretera Sacramento s/n, 04120, Almería, Spain. vjara@ual.es
Cámara-Artigas
A
A
Jara-Pérez
V
V
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D008168:Lung / Q000201:enzymology*; D008970:Molecular Weight; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet; D013552:Swine
D007703:Peptidyl-Dipeptidase A