Secretion of triacylglycerol-poor VLDL particles from McA-RH7777 cells expressing human hepatic lipase |
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Authors: | Bamji-Mirza Michelle Sundaram Meenakshi Zhong Shumei Yao Erik F Parks Robin J Yao Zemin |
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Institution: | Department of Biochemistry, Microbiology & Immunology, Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Canada, K1H 8M5. |
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Abstract: | Hepatic lipase (HL) plays a role in the catabolism of apolipoprotein (apo)B-containing lipoproteins through its lipolytic and ligand-binding properties. We describe a potential intracellular role of HL in the assembly and secretion of VLDL. Transient or stable expression of HL in McA-RH7777 cells resulted in decreased (by 40%) incorporation of (3)H]glycerol into cell-associated and secreted triacylglycerol (TAG) relative to control cells. However, incorporation of (35)S]methionine/cysteine into cell and medium apoB-100 was not decreased by HL expression. The decreased (3)H-TAG synthesis/secretion in HL expressing cells was not attributable to decreased expression of genes involved in lipogenesis. Fractionation of medium revealed that the decreased (3)H]TAG from HL expressing cells was mainly attributable to decreased VLDL. Expression of catalytically-inactive HL (HL(SG)) (Ser-145 at the catalytic site was substituted with Gly) in the cells also resulted in decreased secretion of VLDL-(3)H]TAG. Examination of lumenal contents of microsomes showed a 40% decrease in (3)H]TAG associated with lumenal lipid droplets in HL or HL(SG) expressing cells as compared with control. The microsomal membrane-associated (3)H]TAG was decreased by 50% in HL expressing cells but not in HL(SG) expressing cells. Thus, expression of HL, irrespective of its lipolytic function, impairs formation of VLDL precursor (3)H]TAG in the form of lumenal lipid droplets. These results suggest that HL expression in McA-RH7777 cells result in secretion of (3)H]TAG-poor VLDL. |
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Keywords: | apolipoprotein B-100 heparin sulfate proteoglycans lumenal lipid droplet microsomal triglyceride transfer protein triacylglycerol very low density lipoprotein |
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