Isolation and properties of rat plasma lecithin-cholesterol acyltransferase.
Lecithin-cholesterol acyltransferase was purified from rat plasma and the properties of this enzyme during the purification procedures and those of the purified enzyme were investigated in comparison with the human enzyme. The rat enzyme was not adsorbed on hydroxyapatite, which was employed for the purification of the human enzyme. When purified human enzyme was incubated at 37 degrees C in 0.1 mM phosphate buffer (pH 7.4; ionic strength, 0.00025), no alteration of enzyme activity was observed for up to 6 h. In the case of the rat enzyme, however, approximately 40% of the enzyme activity was lost under the same conditions. The human enzyme and rat enzyme were both retained on a Sepharose 4B column to which HDL3 was covalently linked, in 39 mM phosphate buffer, pH 7.4. Although the human enzyme was eluted from the column in 1 mM phosphate buffer, the rat enzyme was dissociated from the column at a lower buffer concentration (0.1 mM phosphate buffer). These findings indicate that the rat enzyme effectively associated with HDL3 in 39 mM phosphate buffer, pH 7.4, but the association was more sensitive to increase of ionic strength compared with that of the human enzyme.
Furukawa
Y
Y
Department of Food Chemistry, Faculty of Agriculture, Tohoku University, Miyagi.
Urano
T
T
Itoh
H
H
Takahashi
C
C
Kimura
S
S
D000818:Animals; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000032:analysis / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D003911:Dextrans / Q000494:pharmacology; D006801:Humans; D008075:Lipoproteins, HDL / Q000032:analysis; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000032:analysis / Q000097:blood* / Q000302:isolation & purification; D051381:Rats; D012710:Serum Albumin, Bovine / Q000032:analysis; D013045:Species Specificity; D014461:Ultracentrifugation
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D003911:Dextrans; D008075:Lipoproteins, HDL; D012710:Serum Albumin, Bovine; D007862:Phosphatidylcholine-Sterol O-Acyltransferase