LIPOPROTEINS All lipoproteins are soluble, spherical lipid traffickers which consist of a hydrophobic triglyceride (TG) and CE core,

surrounded by a surface layer of free cholesterol (FC), phospholipids and apolipoprotiens. Lipoprotein types are classified based on their density, lipid composition and association with apolipoproteins. The largest lipoproteins, chylomicrons, are formed in the intestine and transport dietary TG on an apolipoprotein B48 (apoB48) scaffold. They are also associated with apolipoproteins Al, All, AV, C and E. Very low density lipoproteins (VLDL) transport endogenous TG with a modest amount of CE on an apoBIOC scaffold and interact with apoEs and apoCs. VLDL undergoes a series of modifications and lipid exchanges to form CE-enriched LDL particles.

LIPOPROTEIN METABOLISM Whole body lipid homeostasis is maintained through a balance between exogenous cholesterol and fatty acid intake and endogenous synthesis.

EXOGENOUS

LIPOPROTEIN

METABOLISM

Cholesterol absorption is mediated by the transporter Niemann-Pick C1-like1 (NPC1L1). Approximately 50% of dietary cholesterol is absorbed by the intestine. Dietary fat is hydrolyzed by pancreatic lipase, absorbed very efficiently in the intestine and dictates the rate of formation of TG-rich chylomicrons . The protein scaffold for assembly of cholesterol and TG into chylomicron particles is apolipoprotein B. ApoB exists in two isoforms, which arise from editing of the APOB mRNA transcript. ApoB48 is the N-terminal 48% of the 550 kDa polypeptide chain which is edited by apoB-encoding mRNA (apobec). In the human intestine, apoB48 is the main isoform, while full length apoB100 is expressed exclusively in human liver. In mice, both the liver

and intestine produce apoB48. TG and cholesterol, in addition to PL are packaged onto the apoB48 protein backbone by microsomal triglyceride transport protein (MTP) within intestinal enterocytes prior to secretion into the lymphatic system . Once chylomicrons enter the bloodstream, they become associated with apolipoproteins E and C. They then bind heparin sulfate proteoglycans, and apoCII is required for TG hydrolysis by lipoprotein lipase, an enzyme which is secreted by parenchymal cells of muscle and adipose and resides on the capillary endothelium. Released fatty acids and glycerol diffuse through the capillary walls to be taken up and utilized or reesterified and stored by peripheral tissues.CE rich remnant particles are taken up very efficiently by the liver through LDLR- and LDL related receptor (LRP)- mediated recognition of apoE

ENDOGENOUS LIPOPROTEIN METABOLISM Since only a small component of circulating cholesterol is derived from the diet, the liver is the primary regulator of whole body cholesterol and lipid metabolism (Hegele, 2009). The liver is composed of hepatocytes, which perform the bulk of liver functions and non-parenchyma! cells including endothelial cells, Kupffer cells and other immune cells. Endothelial cells of capillaries in the liver allow free diffusion of small molecules between the blood and hepatocytes while filtering out larger molecules like chylomicrons, and thus aid in the regulation of lipid uptake by the liver (Bouwens et al., 1992).

Endogenous

and

Exogenous

Lipoprotein

Metabolism

Dietary triglycerides and cholesterol are packaged onto the apolipoprotein B48 scaffold and secreted as chylomicrons, initially into the lymphatics, and subsequently into the blood stream. The lipolysis of triglycerides by lipoprotein lipase (LPL) allows fatty acid uptake into peripheral tissues. The cholesteryl esterrich

remnant particle is taken up by the liver through LDLR-mediated endocytosis. The liver secretes TG-rich VLDL particles, which also undergo hydrolysis by lipoprotein lipase. The subsequent CE-enriched particles, IDL and LDL may also be cleared by the liver through the LDLR. Cholesterol from peripheral tissues is effluxed to HDL and which is also taken up by the liver through SRB1 and secreted into bile. In addition to the uptake of dietary cholesterol and fatty acids, the endogenous synthesis of both lipids also occurs in the liver. Both cholesterol and triglyceride synthesis begin with the formation of acetyl-CoA, derived from the metabolism of glucose or fatty acids. Cholesterol synthesis is very complex, involving greater than 30 individual reactions. It requires the condensation of two acetyl-CoA molecules to create acetoacetyl-CoA which is subsequently converted to mevalonate by HMG-CoA reductase, the enzyme target of statin drugs. Through a series of condensation reactions, isoprene units are formed which condense to form squalene. Squalene is cyclized to form lanosterol, which is subsequently converted to cholesterol through a further 19 step process. The synthesis of fatty acids begins when acetyl-CoA is carboxylated to form malonyl-CoA, which is subsequently elongated to palmitic acid by fatty acid synthase (FAS). Both cholesterol and triglyceride availability within the liver have been shown to regulate lipoprotein secretion. Assembly of the TG-rich core onto the apoB100 backbone happens exclusively in the liver and VLDL is the primary lipoprotein synthesized and secreted. Within the bloodstream, lipoprotein lipase (LPL) initiates lipolysis of TG within VLDL, resulting in the conversion of VLDL into intermediate-density lipoprotein (IDL), and fatty acid uptake via CD36 by peripheral tissues such as adipose and muscle. Once the VLDL are reduced in size, they can become enriched in apoE, which leads to uptake via recognition of apoE

by the LDLR and LDLR related protein (LRP) (Hu et al., 2008). IDL can also be converted into LDL by further lipolysis and the addition of CE through the activity of CETP. These TG-enriched LDL particles are substrates for hepatic lipase, which further reduces particle size, resulting in small, dense LDL particles. Lipoprotein and fatty acid uptake can play a significant role in regulating lipid levels within tissues. The LDLR, which binds the apoB backbone of LDL, is expressed 19 ubiquitously and can mediate peripheral lipoprotein uptake. However both the LDLR and the LDLR related protein (LRP) are expressed predominantly in liver, which mediates the clearance of most lipoprotein particles from plasma (Espirito Santo et al., 2005).

HEPATIC LIPOPROTEIN SYNTHESIS In order to be successfully secreted into plasma, apoB must be transcribed, fully translated and the polypeptide translocated, lipidated, properly folded and glycosylated. Thus the complexity of this process allows for multiple points of regulation during apoB particle synthesis and assembly