Interaction of apolipoprotein(a) with apolipoprotein B-containing lipoproteins.
Recombinant DNA-derived apolipoprotein(a) was used to demonstrate that the apo(a) moiety of lipoprotein(a) (Lp(a)) is responsible for the binding of Lp(a) to other apolipoprotein B-containing lipoproteins (apoB-Lp) including LDL2, a subclass of low density lipoproteins (d = 1.030-1.063 g/ml). The r-apo(a).LDL2 complexes exhibited the same binding constant as Lp(a).LDL2 (10(-8) M). Treatment of either recombinant apo(a) or Lp(a) with a reducing agent destroyed binding activity. A synthetic polypeptide corresponding to a portion of apo(a)'s kringle-4 inhibited the binding (K1 = 1.9 x 10(-4) M) of LDL2 to Lp(a). Therefore, we concluded that binding to apoB-Lp was mediated by the kringle-4-like domains on apo(a). Using ligand chromatography which can detect complexes having a KD as low as 10(-2) M, we demonstrated the binding of plasminogen to apoB-Lp. Like Lp(a), binding of plasminogen to apoB-Lp was mediated by the kringle domain(s). The differences in binding affinity may be due to amino acid substitutions in the kringle-4-like domain. In most of the kringle-4-like domains of apo(a), the aspartic residue critical for binding to lysine was substituted by valine. Consistent with this substitution, we found that L-proline and hydroxyproline, but not L-lysine, inhibited the binding of LDL2 to apo(a). Inhibition by L-proline could be reversed in the binding studies by increasing the amount of apo(a); and L-proline-Sepharose bound plasma Lp(a), suggesting that L-proline acted as a ligand for the kringle-4-like domain(s) of apo(a) involved in the binding of apoB-Lp. The binding of apo(a) to proline and hydroxyproline could be responsible for the binding of apo(a) to the subendothelial extracellular matrix, i.e. domains of proteins rich in proline or hydroxyproline (e.g. collagen and elastin).
Trieu
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Lipoprotein and Atherosclerosis Research Program, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Zioncheck
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Lawn
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McConathy
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D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001053:Apolipoproteins / Q000378:metabolism*; D001055:Apolipoproteins B / Q000378:metabolism*; D053241:Apoprotein(a); D001667:Binding, Competitive; D015153:Blotting, Western; D002417:Cattle; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel; D005779:Immunodiffusion; D017270:Lipoprotein(a); D008074:Lipoproteins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
D000596:Amino Acids; D001053:Apolipoproteins; D001055:Apolipoproteins B; D017270:Lipoprotein(a); D008074:Lipoproteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D010958:Plasminogen; D053241:Apoprotein(a); D004229:Dithiothreitol