Isolation and characterization of the two major apoproteins in human lipoprotein [a].
Human Lp[a] was isolated in preparative amounts from two donors; the native lipoprotein and its constituent apoproteins, apo[a] and apoB, were characterized extensively. Based on differences in apparent molecular weight, four different isoforms of apo[a], a1-a4, were observed between the two donors. The number and relative distribution of these isoforms varied between donors but were constant for each donor. Each apo[a] isoform was shown to be derived from a discrete apo[a]-B100 disulfide-linked complex present before reduction. Complete delipidation of Lp[a] was followed by solubilization, reduction, and carboxamidomethylation of the constituent apoproteins. These apoproteins were then separated by immunoaffinity chromatography using anti-apo[a]- or anti-apoB-Sepharose; their purity and structural integrity were demonstrated by Western blot analysis. ApoB isolated by this procedure was essentially identical to apoB from autologous LDL with respect to molecular weight, secondary structure, amino acid composition, and sialic acid content. However, apo[a] differed from apoB in that it exhibited: a much less alpha-helical, less beta, but much more disordered structure; a lower proportion of aspartate, isoleucine, leucine, phenylalanine, and lysine, but a higher proportion of proline, glycine, and threonine; and a much higher content of sialic acid. These results indicate that apo[a] is not a superglycosylated form of apoB but is distinctly different in its composition and structure.
D000596:Amino Acids / Q000032:analysis; D001053:Apolipoproteins / Q000097:blood* / Q000302:isolation & purification; D053241:Apoprotein(a); D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007122:Immunoelectrophoresis; D017270:Lipoprotein(a); D008074:Lipoproteins / Q000097:blood* / Q000302:isolation & purification; D008077:Lipoproteins, LDL / Q000097:blood; D012794:Sialic Acids / Q000032:analysis; D012995:Solubility
D000596:Amino Acids; D001053:Apolipoproteins; D017270:Lipoprotein(a); D008074:Lipoproteins; D008077:Lipoproteins, LDL; D012794:Sialic Acids; D053241:Apoprotein(a)