Fatty acids bound to human serum albumin and its structural variants modulate apolipoprotein B secretion in HepG2 cells

• Published in: Biochimica et biophysica acta Vol. 1761; no. 7; pp. 717 - 724
• Main Authors: Ha, Ji-Sook, Theriault, Andre, Bhagavan, Nadhipuram V., Ha, Chung-Eun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2006
• Subjects: Apolipoprotein B100; Apolipoproteins B - secretion; Cell Line, Tumor; Coronary Disease - etiology; Coronary Disease - metabolism; Fatty acid; Hepatocytes - metabolism; HepG2 cell; Human serum albumin; Humans; In vitro protein expression; Mutagenesis, Site-Directed; Mutation; Oleic Acid - metabolism; Protein Binding; Serum Albumin - chemistry; Serum Albumin - genetics; Serum Albumin - metabolism; Site-directed mutagenesis; Structure-Activity Relationship; Human serum albumin; Apolipoprotein B100; In vitro protein expression; HepG2 cell; Site-directed mutagenesis; Fatty acid
• DOI: 10.1016/j.bbalip.2006.05.016
• ISSN: 1388-1981; 0006-3002

Epidemiologic studies have shown an inverse relationship between human serum albumin (HSA) levels and coronary heart disease (CHD). However, no mechanisms have been identified to explain this relationship. We hypothesized that this relationship is due to differences in binding affinity of fatty acids to HSA and subsequent atherogenic lipoprotein synthesis and secretion from hepatocytes. To test our hypothesis we undertook the current study. Using HepG2 cells, we demonstrated that oleic acid (OA) bound to HSA in a molar ratio of 4:1 and after incubation for 24 h stimulated apolipoprotein B (apoB) secretion. We also tested whether mutant forms of HSA could alter the binding affinity for fatty acids and change the availability of substrate for lipoprotein secretion. Based on the results obtained in this study using 11 HSA mutant proteins complexed with OA, we were able to classify into three major mutant groups based on their effects on apoB secretion. One group in particular (R410Q/Y411W, R410A/Y411A, and W214L/Y411W) showed a significantly diminished effect on apoB secretion when compared to the wild type HSA/OA complex. Furthermore, the amount of free OA internalized in HepG2 cells in the presence of HSA mutant proteins was in good agreement with the effects seen on apoB secretion by the various HSA mutants. This suggests that some mutant forms of HSA might potentially bind fatty acids with a much higher binding affinity and thus deprive fatty acids available for lipoprotein assembly in hepatocytes. In conclusion, our data illustrate that certain HSA polymorphic forms may be protective against the development of CHD and warrants further investigation.