Oxidation-labile subfraction of human plasma low density lipoprotein isolated by ion-exchange chromatography.
We isolated subfractions of human plasma low density lipoprotein (LDL) using ion-exchange chromatography. Plasma LDL from normolipidemic subjects were applied to a DEAE Sepharose 6B column. After elution of the bulk of LDL at 150 mM NaCl (the major fraction), the residual LDL was eluted at 500 mM NaCl and designated as the minor fraction. The minor fraction, only less than 1% of total LDL, tended to be somewhat similar in certain properties to oxidized LDL, e.g., an increased negative charge, higher protein/cholesterol ratio, and a higher flotation density than native LDL. These results were consistent with data reported by Avogaro et al. (1988. Arteriosclerosis. 8: 79-87). However, assays of 125I-labeled LDL binding activity for LDL receptors equal to that of the major fraction. Incorporation of [14C]oleate into cholesteryl ester [acyl-CoA:cholesterol acyltransferase (ACAT) activity] in mouse peritoneal macrophages incubated with the minor fraction was only slightly greater than that with the major fraction. Incubation of the minor fraction with 0.5 microM Cu2+ caused a remarkable stimulation of ACAT activity, while stimulation by the major fraction required incubation with 5 microM Cu2+, suggesting that the minor fraction was relatively labile to oxidation. The minor but definite presence of a plasma LDL subfraction more negative and susceptible to oxidation implicates the possibility of its association with atherogenesis.
Shimano
H
H
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Yamada
N
N
Ishibashi
S
S
Mokuno
H
H
Mori
N
N
Gotoda
T
T
Harada
K
K
Akanuma
Y
Y
Murase
T
T
Yazaki
Y
Y
D001161:Arteriosclerosis / Q000209:etiology; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D003300:Copper / Q000378:metabolism; D004563:Electrochemistry; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008077:Lipoproteins, LDL / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D010084:Oxidation-Reduction; D011973:Receptors, LDL / Q000378:metabolism
D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D003300:Copper