Analysis of the mechanism of lipoprotein(a) assembly.
We have assessed the ability of a battery of purified recombinant apolipoprotein(a) (r-apo(a)) derivatives to bind to immobilized low-density lipoprotein (LDL) by ELISA. Removal of the apo(a) kringle IV type 8 and type 9 sequences dramatically reduced apo(a) binding to LDL. The binding of apo(a) to LDL was effectively inhibited by arginine, lysine, the lysine analogue epsilon-aminocaproic acid and proline; comparable inhibition was observed using the 17K and KIV5-8 r-apo(a) derivatives, suggesting a direct role for sequences contained in the latter species in mediating the initial non-covalent interactions which precede specific disulfide bond formation. We also determined that r-apo(a) binds directly to a synthetic apoB peptide spanning amino acid residues 3732-3745; this interaction appeared to be mediated by sequences present in apo(a) kringle IV types 8 and 9, and could be inhibited by arginine, lysine and proline. The results of this study indicate that the efficiency of Lp(a) assembly is a direct function of the initial non-covalent interactions between apo(a) and LDL; in addition, these studies suggest that Cys3734 in apoB mediates covalent linkage with apo(a) by virtue of the ability of the apoB sequences surrounding this residue to directly interact with apo(a) KIV type 9.
Koschinsky
M L
ML
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada. MK11@post.queensu.ca
Marcovina
S M
SM
May
L F
LF
Gabel
B R
BR
D001054:Apolipoproteins A / Q000378:metabolism*; D001055:Apolipoproteins B / Q000737:chemistry / Q000378:metabolism; D001120:Arginine / Q000494:pharmacology; D002460:Cell Line; D006801:Humans; D018082:Kringles / Q000502:physiology; D017270:Lipoprotein(a) / Q000096:biosynthesis*; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D011392:Proline / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D011994:Recombinant Proteins / Q000378:metabolism
D001054:Apolipoproteins A; D001055:Apolipoproteins B; D017270:Lipoprotein(a); D008077:Lipoproteins, LDL; D010446:Peptide Fragments; D011994:Recombinant Proteins; D010958:Plasminogen; D001120:Arginine; D011392:Proline; D008239:Lysine