title: Isolation of angiotensin converting enzyme (ACE) binding protein from human serum with an ACE affinity column.
abstract: Immobilized angiotensin-converting enzyme (ACE) was utilized as an affinity ligand to isolate a naturally occurring ACE binding protein from normal human serum. The enzyme was isolated from solubilized bovine lung membrane preparations by lisinopril affinity chromatography. It had an estimated molecular weight of 180 000 and was recognized by the anti-ACE antibody for the rabbit testicular ACE in immunoblots. ACE was immobilized onto epoxy Sepharose as well as Affi-Gel 15. Immobilized ACE on Affi-Gel 15 had higher catalytic activity (0.176 U/mL) compared with the enzyme immobilized on epoxy Sepharose (0.00005 U/mL). Immobilized ACE served as the affinity ligand for the identification of the ACE binding protein in human serum with an estimated molecular weight of 14 000 as observed by SDS polyacrylamide gel electrophoresis. The identification and further characterization of ACE binding proteins in serum and tissues may facilitate the greater understanding of the endogenous regulation of this key enzyme, which is involved in blood pressure homeostasis.
authors.affiliation: Department of Chemistry, Bowling Green State University, OH 43403, USA.
authors.forename: S
authors.initials: S
authors.lastname: Thevananther
authors.forename: A S
authors.initials: AS
authors.lastname: Brecher
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000097:blood / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity; D004800:Enzymes, Immobilized; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D017706:Lisinopril / Q000378:metabolism; D008970:Molecular Weight; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D011817:Rabbits
chemical_list: D002352:Carrier Proteins; D004800:Enzymes, Immobilized; D017706:Lisinopril; D007703:Peptidyl-Dipeptidase A