TY - JOUR
T1 - Characterization of very-low-density lipoproteins isolated from baboons, and fractionated using heparin-Sepharose chromatography
AU - Rainwater, David L.
AU - Kushwaha, Rampratap S.
N1 - Funding Information:
This work was supported by grants from the National Institutes of Health (HL-28972 and NHLBI Contract No. 53030) the Andrew G. Cowles Research Fellowship, and a gift from Mrs. Betty Roberts Kelso. The authors would like to thank Drs. Anthony M. Coelho, Jr. and K.D. Carey for the baboon blood and Glen E. Mott for the apolipoprotein B measurements. In addition, we thank Mary Huffman, Mary Williams, Mary Sparks, Scott Manis and Judy Odom for their assistance in these studies.
PY - 1988/2/19
Y1 - 1988/2/19
N2 - Plasma very-low-density lipoproteins (VLDL) (d< 1.006 g/ml) were purified from baboons by repeated ultracentrifugation. The weight composition of VLDL purified from these animals was 59% triacylglycerol, 17% phospholipid, 13% cholesterol plus cholesteryl esters, and 11% protein. When purified VLDL was fractionated using heparin-Sepharose chromatography, an average of 33% of the total recovered proteins were unbound in a saline solution, and 67% (range, 31 to 92%) were bound by the column, but could be eluted with 3 M NaCl. Recoveries of starting protein and the major classes of lipids in the two fractions were 70-80%. The two fractions differed in both apolipoprotein and lipid compositions. Analysis of sodium dodecyl sulfate-treated apolipoproteins using 3-21.5% acrylamide gradient gel electrophoresis indicated that both VLDL fractions contained apolipoprotein B, but only the bound fraction possessed significant amounts of apolipoprotein E. On a weight percent basis, the apolipoprotein-E-rich (bound) VLDL fraction contained significantly more cholesterol and cholesteryl esters (P < 0.001) and less phospholipids (P < 0.005) compared to the apolipoprotein E-poor (unbound) VLDL. Apolipoprotein-E-poor VLDL had shorter retention times than E-rich VLDL upon gel filtration chromatography, suggesting a larger size. There was no significant correlation between plasma levels of apolipoprotein-E-poor VLDL and levels of apolipoprotein B. These results demonstrate that baboons possess VLDL which can be separated into apolipoprotein-E-poor and E-rich fractions and these fractions differ in protein and lipid composition and in size.
AB - Plasma very-low-density lipoproteins (VLDL) (d< 1.006 g/ml) were purified from baboons by repeated ultracentrifugation. The weight composition of VLDL purified from these animals was 59% triacylglycerol, 17% phospholipid, 13% cholesterol plus cholesteryl esters, and 11% protein. When purified VLDL was fractionated using heparin-Sepharose chromatography, an average of 33% of the total recovered proteins were unbound in a saline solution, and 67% (range, 31 to 92%) were bound by the column, but could be eluted with 3 M NaCl. Recoveries of starting protein and the major classes of lipids in the two fractions were 70-80%. The two fractions differed in both apolipoprotein and lipid compositions. Analysis of sodium dodecyl sulfate-treated apolipoproteins using 3-21.5% acrylamide gradient gel electrophoresis indicated that both VLDL fractions contained apolipoprotein B, but only the bound fraction possessed significant amounts of apolipoprotein E. On a weight percent basis, the apolipoprotein-E-rich (bound) VLDL fraction contained significantly more cholesterol and cholesteryl esters (P < 0.001) and less phospholipids (P < 0.005) compared to the apolipoprotein E-poor (unbound) VLDL. Apolipoprotein-E-poor VLDL had shorter retention times than E-rich VLDL upon gel filtration chromatography, suggesting a larger size. There was no significant correlation between plasma levels of apolipoprotein-E-poor VLDL and levels of apolipoprotein B. These results demonstrate that baboons possess VLDL which can be separated into apolipoprotein-E-poor and E-rich fractions and these fractions differ in protein and lipid composition and in size.
KW - (Baboon)
KW - Apolipoprotein E
KW - Heparin-Sepharose
KW - Lipoprotein
KW - VLDL
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U2 - 10.1016/0005-2760(88)90227-5
DO - 10.1016/0005-2760(88)90227-5
M3 - Article
C2 - 3342249
AN - SCOPUS:0024291475
VL - 958
SP - 416
EP - 423
JO - BBA - Specialised Section On Lipids and Related Subjects
JF - BBA - Specialised Section On Lipids and Related Subjects
SN - 1388-1981
IS - 3
ER -