Molecular requirements in the recognition of low-density lipoproteins (LDL) by specific platelet membrane receptors.
We have demonstrated that platelet low-density lipoprotein (LDL) receptors differ from classic LDL receptors of nucleated cells. Although positively charged Arg and Lys residues of apoprotein B-100 are known to play a key role in LDL recognition by classic LDL receptors, there are no conclusive data on platelet LDL receptors. This study investigated the molecular requirements of LDL particle recognition by platelet LDL receptors. The involvement of lipid and protein fractions was determined by displacement studies of the binding of 125I-LDL to platelets and fibroblasts (used as a classical LDL receptor model). The role of the protein moiety was evaluated by chemically modifying positively charged apoB residues (Lys, Arg, and Tyr) via copper-induced oxidation, cyclohexanedione, and tetranitromethane, respectively. The involvement of the lipid fraction was determined by ligand binding assays using 125I-LDL particles that had previously been delipidated and subjected to apoB solubilization. The degree of particle modification was analyzed by agarose/acrylamide gel electrophoresis and anion exchange chromatography. Modifying the amino acid residues increased particle electronegativity in the following order of potency: CHD-LDL>TNM-LDL>ox-LDL>native LDL. The results obtained by displacement studies in fibroblasts suggested that the gain in the LDL negative charge was the most important factor in the loss of receptor affinity. The chemical models of protein modification used in our study greatly affected LDL binding to the classical fibroblast receptor. In contrast, there was very slight difference in the displacement capacity on platelet 125I-LDL binding, which suggests that the protein fraction does not play a major role in the interaction of LDL with its platelet receptor. On the other hand, whereas modifying the lipid moiety did not alter the ability of solubilized 125I-apoB to interact with the classical fibroblast LDL receptor, platelet LDL receptors were unable to recognize these particles. In conclusion, our results confirm that the protein fraction plays a key role in the fibroblast LDL-receptor recognition process, whereas the lipid fraction appears to have a more relevant role in platelet LDL-receptor recognition.
Pedreño
J
J
Unitat de Recerca en Lipids i Arteriosclerosi, Universitat Rovira i Virgili, Facultat de Medicina, Reus, Spain. jp@fmcs.urv.es
Sánchez-Quesada
J L
JL
Cabré
A
A
Masana
L
L
D001055:Apolipoproteins B / Q000737:chemistry / Q000378:metabolism; D001120:Arginine / Q000737:chemistry; D001665:Binding Sites; D001667:Binding, Competitive; D001792:Blood Platelets / Q000737:chemistry / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D004587:Electrophoresis, Agar Gel; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes; D008024:Ligands; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry / Q000502:physiology; D008077:Lipoproteins, LDL / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000378:metabolism; D011973:Receptors, LDL / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry
D001055:Apolipoproteins B; D007457:Iodine Radioisotopes; D008024:Ligands; D008055:Lipids; D008077:Lipoproteins, LDL; D011956:Receptors, Cell Surface; D011973:Receptors, LDL; D014443:Tyrosine; D001120:Arginine; D008239:Lysine