title: Structural and functional properties of apolipoprotein B in chemically modified low density lipoproteins.
abstract: The structural and compositional changes occurring during in vitro chemical modification of apolipoprotein B-100 (apo B), the apolipoprotein component of low density lipoproteins (LDL), were investigated in this study. The functional properties of chemically modified apo B and especially its potential to induce accumulation of cholesterol esters in macrophages were related to the structural changes of apo B. Acetylation, maleylation or malondialdehyde conjugation did not significantly affect the lipid composition of LDL. However, the unsaturated cholesteryl esters content, especially that of cholesteryl arachidonate was significantly decreased through Cu-oxidation. The number of reactive lysine residues in apo B was decreased by Cu-catalyzed LDL oxidation, acetylation, maleylation and by malondialdehyde conjugation. The number of free cysteines decreased from six in native apo B-100 to three in Cu-oxidized LDL. The tryptophan fluorescence intensity decreased most in malondialdehyde-conjugated LDL and in Cu-oxidized LDL, compared with acetylated and maleylated LDL. The secondary structure of native and chemically modified LDL was measured by attenuated total reflection infrared spectroscopy and by circular dichroism. No significant changes were observed in the secondary structure of any of the modified LDL. These data suggest that neither acetylation, malondialdehyde treatment or even Cu-oxidation substantially altered the secondary structure of apo B, in spite of significant modifications in the primary structure. Incubation of chemically modified LDL with J774 macrophages induced an accumulation of cellular cholesteryl esters and foam cell formation. The highest cholesterol accumulation was induced after malondialdehyde treatment of LDL. These data suggest that the cellular uptake and accumulation of modified LDL is not modulated by changes in the apo B structure. Rather it seems dependent upon the net charge of the apo B protein and probably involves the modification of critical lysine residues.
authors: Dept. Clinical Chemistry, A.Z. St-Jan, Brugge, Belgium.
authors: L
authors: L
authors: Vanderyse
authors: A M
authors: AM
authors: Devreese
authors: J
authors: J
authors: Baert
authors: B
authors: B
authors: Vanloo
authors: L
authors: L
authors: Lins
authors: J M
authors: JM
authors: Ruysschaert
authors: M
authors: M
authors: Rosseneu
mesh_terms: D000107:Acetylation; D000596:Amino Acids / Q000032:analysis; D001055:Apolipoproteins B / Q000737:chemistry* / Q000502:physiology*; D002460:Cell Line; D002788:Cholesterol Esters / Q000032:analysis; D006801:Humans; D008055:Lipids / Q000032:analysis; D008077:Lipoproteins, LDL / Q000737:chemistry* / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008315:Malondialdehyde; D010084:Oxidation-Reduction; D017392:Thiobarbituric Acid Reactive Substances
chemical_list: D000596:Amino Acids; D001055:Apolipoproteins B; D002788:Cholesterol Esters; D008055:Lipids; D008077:Lipoproteins, LDL; D017392:Thiobarbituric Acid Reactive Substances; D008315:Malondialdehyde