Identification of Apo B-100 segments mediating the interaction of low density lipoproteins with arterial proteoglycans.
The interactions of low density lipoprotein (LDL) and apolipoprotein (apo) B-100 segments with chondroitin-6-SO4 rich aortic proteoglycans aggregate (CSPG) were studied by using quantitative frontal elution affinity chromatography. The affinity of the agarose-CSPG was higher for LDL than for very low density lipoprotein and high density lipoprotein was not bound. LDL from different individuals had dissociation coefficients (Kd) from 28 to 179 nM. Experiments with tryptic hydrolysates of apo B suggested that the capacity of LDL to bind with CSPG resides in the protein. Nine apo B-100 hydrophilic peptides, 12 to 26 amino acids long, were selected, and three were found to interact with the agarose-bound CSPG: apo B P-1 (LRKHKLIDVISMYRELLKDLSKEA, residues 4230 to 4254), apo B P-2 (RLTRKRGLKLATALSLSNK, residues 3359 to 3377), and apo B P-11 (RQVSHAKEKLTALTKK, residues 2106 to 2121). These peptides competed with LDL for binding to the agarose-bound and soluble CSPG; apo B P-2 was the most effective. This correlates with Kd values: 63, 86, and 82 microM for apo B P-2, P-1, and P-11, respectively. The peptides shared an excess of positive-charged side chains. Apo B P-2 belongs to the lys- and arg-rich, LDL-receptor domain. Apo E also binds to the agarose-proteoglycan. The results suggest that apo B regions with sequences and charge distributions analogous to those of residues 3359 to 3377, 4230 to 4254, and 2106 and 2121 are among those responsible for the interaction of LDL with intima-media CSPG.
Camejo
G
G
Instituto Venezolano de Investigaciones Cientificas, Centro de Biofisica y Bioquimica, Caracas, Venezuela.
Olofsson
S O
SO
Lopez
F
F
Carlsson
P
P
Bondjers
G
G
D001011:Aorta / Q000378:metabolism*; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000138:chemical synthesis / Q000494:pharmacology*; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D002846:Chromatography, Affinity; D006801:Humans; D066298:In Vitro Techniques; D008077:Lipoproteins, LDL / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism*
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D011508:Chondroitin Sulfate Proteoglycans; D008077:Lipoproteins, LDL; D010446:Peptide Fragments; D011509:Proteoglycans