title: Partial purification of phosphoramidon-sensitive endothelin converting enzyme in porcine aortic endothelial cells: high affinity for Ricinus communis agglutinin.
abstract: A membrane-bound endothelin converting enzyme (ECE) of porcine aortic endothelial cells (ECs) was solubilized with Lubrol PX with high efficiency and stability. The solubilized ECE was bound to Ricinus communis agglutinin (RCA) but not to peanut agglutinin (PNA) or wheat germ agglutinin (WGA), suggesting that the ECE has a galactosylated structure possessing a high affinity for RCA. The sequential chromatography on RCA-agarose, PNA-agarose and a TSKgel DEAE-5PW column attained 2,100-fold purification for the ECE over the membrane fractions. The purified ECE was sensitively inhibited by phosphoramidon but not by thiorphan. The present RCA and PNA affinity column procedures may be a powerful approach to isolation of ECE of EC origin.
authors.affiliation: Third Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ohnaka
authors.forename: M
authors.initials: M
authors.lastname: Nishikawa
authors.forename: R
authors.initials: R
authors.lastname: Takayanagi
authors.forename: M
authors.initials: M
authors.lastname: Haji
authors.forename: H
authors.initials: H
authors.lastname: Nawata
mesh_terms: D000818:Animals; D001011:Aorta / Q000201:enzymology; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D000072396:Endothelin-Converting Enzymes; D004730:Endothelium, Vascular / Q000201:enzymology*; D006020:Glycopeptides / Q000494:pharmacology; D037102:Lectins / Q000378:metabolism; D008666:Metalloendopeptidases; D008970:Molecular Weight; D019887:Peanut Agglutinin; D037121:Plant Lectins; D012995:Solubility; D013552:Swine
chemical_list: D006020:Glycopeptides; D037102:Lectins; D019887:Peanut Agglutinin; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes; C008890:phosphoramidon