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Purification of angiotensin I converting enzyme from pig lung using concanavalin-A sepharose chromatography
M Andujar-Sánchez1, A Cámara-Artigas, V Jara-Pérez
1Dpto. Química Física, Bioquímica y Qui;mica Inorgánica, Universidad de Almería, Carretera Sacramento s/n, 04120, Almería, Spain. vjara@ual.es
Abstract:
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.