Epitopes close to the apolipoprotein B low density lipoprotein receptor-binding site are modified by advanced glycation end products.
Advanced glycation end products (AGEs) are thought to contribute to the abnormal lipoprotein profiles and increased risk of cardiovascular disease of patients with diabetes and renal failure, in part by preventing apolipoprotein B (apoB)-mediated cellular uptake of low density lipoproteins (LDL) by LDL receptors (LDLr). It has been proposed that AGE modification at one site in apoB, almost 1,800 residues from the putative apoB LDLr-binding domain, may be sufficient to induce an apoB conformational change that prevents binding to the LDLr. To further explore this hypothesis, we used 29 anti-human apoB mAbs to identify other potential sites on apoB that may be modified by in vitro advanced glycation of LDL. Glycation of LDL caused a time-dependent decrease in its ability to bind to the LDLr and in the immunoreactivity of six distinct apoB epitopes, including two that flank the apoB LDLr-binding domain. ApoB appears to be modified at multiple sites by these criteria, as the loss of glycation-sensitive epitopes was detected on both native glycated LDL and denatured, delipidated glycated apoB. Moreover, residues directly within the putative apoB LDLr-binding site are not apparently modified in glycated LDL. We propose that the inability of LDL modified by AGEs to bind to the LDLr is caused by modification of residues adjacent to the putative LDLr-binding site that were undetected by previous immunochemical studies. AGE modification either eliminates the direct participation of the residues in LDLr binding or indirectly alters the conformation of the apoB LDLr-binding site.
Wang
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Lipoprotein and Atherosclerosis Group, University of Ottawa Heart Institute, Ottawa, ON, Canada K1Y 4E9.
Bucala
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Milne
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D000911:Antibodies, Monoclonal / Q000378:metabolism; D001055:Apolipoproteins B / Q000276:immunology / Q000378:metabolism*; D001665:Binding Sites / Q000276:immunology; D001667:Binding, Competitive; D002460:Cell Line; D000939:Epitopes / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006801:Humans; D011863:Radioimmunoassay; D011973:Receptors, LDL / Q000276:immunology / Q000378:metabolism*; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D000939:Epitopes; D017127:Glycation End Products, Advanced; D011973:Receptors, LDL