Isolation and partial characterization of apolipoprotein (a) from human lipoprotein (a)

• Published in: Biochemistry and cell biology Vol. 64; no. 10; p. 999
• Main Authors: Seman, L J, Breckenridge, W C
• Format: Journal Article
• Language: English
• Published: Canada 01.10.1986
• Subjects: Amino Acids - analysis; Apolipoproteins A - blood; Apolipoproteins A - isolation & purification; Apolipoproteins B - blood; Apolipoproteins B - isolation & purification; Carbohydrates - analysis; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Humans; Lipids - analysis; Lipoprotein(a); Lipoproteins - blood
• ISSN: 0829-8211
• DOI: 10.1139/o86-133

A procedure was developed for the dissociation of apolipoprotein (a) (apo (a)) from pure human lipoprotein (a) (Lp(a)) prepared by density gradient ultracentrifugation and gel filtration. Lp(a) was ultracentrifuged through a layer of saline which was adjusted to a density of 1.182 g/mL and contained 30 mM dithiothreitol (50 mM) and phenylmethylsulfonyl fluoride (1.25 mM). Following centrifugation, the lipid and apolipoprotein B (apo B) were recovered as a lipoprotein (Lp(a) B) in the supernatant fraction, while the apo (a) was recovered as a lipid-poor protein pellet. An investigation of the supernatant lipoprotein by electron microscopy and compositional analysis revealed that it was similar in size and composition to low density lipoprotein (LDL) isolated from the same density range and contained apo B100 with an amino acid and carbohydrate composition which was similar to apo B from LDL. Estimates of the apparent molecular weight of the apo (a) varied amongst individuals but was always greater than apo B100 (congruent to 450,000). The amino acid composition of apo (a), which was very distinct from apo B, was characterized by a higher content of serine, threonine, proline, and tyrosine, but lower amounts of isoleucine, phenylalanine, and lysine when compared with apo B of Lp(a) or LDL. The apo (a) contained a much higher proportion of carbohydrate, in particular N-acetylgalactosamine, galactose, and N-acetylneuraminic acid (which were three- to six-fold higher) than the apo B of Lp(a). It is concluded that apo (a) is distinct from other apolipoproteins owing to its low avidity for lipid and the nature of the interaction with apo B. Lp(a) consists of an LDL-like particle with a carbohydrate-rich apo (a) attached to the surface of apo B.