Human serum albumin and its structural variants mediate cholesterol efflux from cultured endothelial cells.
In the present study, we used the human EA.hy926 endothelial cell line as the model system to investigate the effect of human serum albumin (HSA) and its structural variants on cholesterol efflux. Initial studies showed that HSA promoted cholesterol efflux in a dose- and time-dependent manner, reaching a plateau at 10 mg/ml at 90 min. As a control, gelatin displayed no significant effect on efflux, while HSA was significantly more efficient than ovalbumin and bovine serum albumin (BSA) in promoting cholesterol efflux. Equal molar concentrations of HSA and apolipoprotein A-I (apoA-I) showed that apoA-I had considerably higher efficiency in efflux. However, the prevailing high plasma concentrations of HSA may compensate for its lower efflux rate compared to apoA-I. To characterize the mechanism of HSA-mediated cholesterol efflux, we studied the effects of cAMP and temperature on efflux using both EA.hy926 endothelial cells and murine RAW 264.7 macrophages. We found that HSA-mediated efflux occurred via a cAMP-independent and relatively temperature-insensitive pathway. We next examined the nature of HSA-cholesterol interaction by comparing the effects of various HSA mutants to wild-type HSA on cholesterol efflux. We found specific interactions between subdomains 2A and 3A and cholesterol, as indicated by the changes in the efflux rate of various HSA mutants. In conclusion, our study provides evidence for the role of HSA in cholesterol efflux, and shows that the substitution of specific amino acid residues in subdomains of 2A and 3A may be important structural determinants in its ability to bind to cholesterol and participate in cholesterol efflux.
Department of Biochemistry and Biophysics, John A. Burns School of Medicine, University of Hawaii at Manoa, 1960 East-West Road, Honolulu, HI 96822, USA.
Ji-Sook
JS
Ha
Chung-Eun
CE
Ha
Jun-Tzu
JT
Chao
Charles E
CE
Petersen
Andre
A
Theriault
Nadhipuram V
NV
Bhagavan
D016632:Apolipoprotein A-I / Q000502:physiology; D001665:Binding Sites; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D000242:Cyclic AMP / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013696:Temperature; D013997:Time Factors
D016632:Apolipoprotein A-I; D012709:Serum Albumin; D002784:Cholesterol; D000242:Cyclic AMP