A new method for isolating the nonidentical protein subunits of human plasma α-lipoprotein

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Abstract

Human plasma alpha lipoprotein (αLP) was totally delipidated by gel filtration on Sephadex LH-20 in a medium of 2 butanol:acetic acid:H2O, 4:1:5. The resulting alpha protein (αP) exhibited two major bands, labeled C and D, on acrylamide-gel electrophoresis in 5.0 M urea at pH 8.8 or 4.0. Minor bands labeled A and B, also present, were shown to be aggregates of C which form when the latter is lyophilized. The C and D components were isolated in pure form from αP (prepared by LH-20 chromatography of αLP) by gel filtration of this protein on Sephadex G-200 in a medium of 1.0 N acetic acid: the C component emerged with a distribution coefficient (Kd) of 0.4, and the D component with a coefficient of 0.7. From each 100 mg of αP, 68 mg of C and 22 mg of D were isolated. 3 mg of a minor fraction with Kd 0.1, containing A and B components as well as C, were also obtained. D but not C reacts with rabbit antiserum to human αLP. C and D differ substantially in content of arginine, histidine, ½-cystine, isoleucine, and tryptophan.

Authors

Daniel Rudman, Luis A. Garcia, Carolyn H. Howard