Conformational changes of apoB-100 in SMase-modified LDL mediate formation of large aggregates at acidic pH.
During atherogenesis, the extracellular pH of atherosclerotic lesions decreases. Here, we examined the effect of low, but physiologically plausible pH on aggregation of modified LDL, one of the key processes in atherogenesis. LDL was treated with SMase, and aggregation of the SMase-treated LDL was followed at pH 5.5-7.5. The lower the pH, the more extensive was the aggregation of identically prelipolyzed LDL particles. At pH 5.5-6.0, the aggregates were much larger (size >1 µm) than those formed at neutral pH (100-200 nm). SMase treatment was found to lead to a dramatic decrease in α-helix and concomitant increase in β-sheet structures of apoB-100. Particle aggregation was caused by interactions between newly exposed segments of apoB-100. LDL-derived lipid microemulsions lacking apoB-100 failed to form large aggregates. SMase-induced LDL aggregation could be blocked by lowering the incubation temperature to 15°C, which also inhibited the changes in the conformation of apoB-100, by proteolytic degradation of apoB-100 after SMase-treatment, and by HDL particles. Taken together, sphingomyelin hydrolysis induces exposure of protease-sensitive sites of apoB-100, whose interactions govern subsequent particle aggregation. The supersized LDL aggregates may contribute to the retention of LDL lipids in acidic areas of atherosclerosis-susceptible sites in the arterial intima.
Wihuri Research Institute, Helsinki,  Finland.
Mia
M
Sneck
Su Duy
SD
Nguyen
Tero
T
Pihlajamaa
Gebrenegus
G
Yohannes
Marja-Liisa
ML
Riekkola
Ross
R
Milne
Petri T
PT
Kovanen
Katariina
K
Oörni
D053299:Apolipoprotein B-100 / Q000737:chemistry* / Q000378:metabolism*; D001409:Bacillus cereus / Q000201:enzymology; D004655:Emulsions; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008066:Lipolysis / Q000187:drug effects; D008077:Lipoproteins, LDL / Q000737:chemistry* / Q000378:metabolism*; D010316:Particle Size; D011485:Protein Binding / Q000187:drug effects; D017433:Protein Structure, Secondary / Q000187:drug effects; D013108:Sphingomyelin Phosphodiesterase / Q000494:pharmacology*
D053299:Apolipoprotein B-100; D004655:Emulsions; D008077:Lipoproteins, LDL; D013108:Sphingomyelin Phosphodiesterase