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Journal of Lipid Research, Vol. 9, 342-349, May 1968
Copyright © 1968 by Lipid Research, Inc.

Properties of human serum low density lipoproteins after modification by succinic anhydride

A. Scanu , H. Pollard , and W. Reader

Departments of Medicine and Biochemistry, The University of Chicago and the Argonne Cancer Research Hospital, Chicago, Illinois 60637

Human serum low density lipoprotein of d 1.019-1.063 (LDL2) treated with succinic anhydride at pH 7.5-8.0 showed the same chemical composition, hydrodynamic properties (flotation and sedimentation coefficients, intrinsic viscosity) and optical properties (circular dichroism) as untreated LDL2. However, in contrast to LDL2, the succinylated product (s-LDL2) failed to react with rabbit anti-LDL2 antisera.

Extraction with ethanol-ether 3:1 yielded the succinylated apoprotein (s-apo-LDL2), which was, unlike untreated apoprotein, soluble in aqueous buffers. Succinylated apoprotein, which was also immunologically unreactive, appeared to differ in structure from s-LDL2, as assessed by the parameters of intrinsic viscosity and circular dichroism.

The molecular weights of both LDL2 and s-LDL2 obtained by the technique of sedimentation equilibrium were 2.1-2.3 x 106. By the same method, s-apo-LDL2 gave an uncorrected figure of 3.95-4.15 x 104 and, after correction for succinyl functions, of 3.60-3.80 x 104. Because of the assumptions made in the computations, the latter figure was considered approximate.

The marked differences in molecular weight between s-apo-LDL2 and whole apo-LDL2 (sim5 x 105) were taken to support the subunit structure of apo-LDL2, which is envisaged as an aggregate of about 12 subunits which dissociate upon succinylation. Further, the large percentage (about 90%) of the free amino groups of LDL2 found to react with succinic anhydride suggests that these groups are at the surface of the molecule.

Supplementary key words serum • low density lipoproteins • succinylated • apoprotein • antigenic reactivity • molecular weight • intrinsic viscosity • circular dichroism • subunit structure

Submitted on December 5, 1967
Accepted on January 23, 1968


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