Intracellular transport of lipids

Summary Translocation of lipids inside mammalian cells is considered to be facilitated by a number of low-molecular weight lipid binding proteins. An overview of these proteins is given, with particular reference to the heart. Three distinct phospholipid transfer proteins specifically stimulate the net transfer of individual phospholipid classes between membrane structures. In rat cardiac muscle their content is 15–140 pmol/g ww. Fatty acid-binding proteins (FABP) are abundantly present in tissues actively involved in the uptake or utilization of long-chain fatty acids, such as intestine, liver and heart. The four distinct FABP types now identified show a complex tissue distribution with some tissues containing more than one type. Heart (H-) FABP comprises about 5% of the cytosolic protein mass; its content in rat heart is 100 nmol/g ww. Immunochemical evidence has been obtained for the presence of H-FABP in several other tissues, including red skeletal muscle, mammary gland and kidney. Beside long-chain fatty acids FABP binds with similar affinity also fatty acyl-CoA and acyl-L-carnitines. In heart the latter compound may be the primary ligand, since normoxic acyl-L-carnitine levels are several fold higher than those of fatty acids. In addition, H-FABP was found to modulate cardiac energy production by controlling the transfer of acyl-L-carnitine to the mitochondrial -oxidative system. H-FABP may also protect the heart against the toxic effects of high intracellular levels of fatty acid intermediates that arise during ischemia.     

Intracellular transport of GPI-anchored proteins

In eukaryotic cells, a subset of proteins are attached to the external leaflet of the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. There is substantial evidence suggesting that these GPI-anchored proteins are clustered in sphingolipid-sterol microdomains or rafts. Since the precursors of these microdomain components are synthesized mainly in the endoplasmic reticulum, it is possible that microdomain assembly occurs during transport along the exocytic route. A sorting mechanism for GPI-anchored proteins using sphingolipid microdomains as selective platforms for vesicle budding has been proposed to operate at different steps in the secretory pathway. Here, we discuss this sorting model in the context of the data obtained from different biological and artificial systems, in addition to other particularities of the intracellular transport of the GPI-anchored proteins.     

Influence of dietary bile acids on formation of bile acids in rat

Various taurine-conjugated bile acids were fed to rats at the 1%-level in the diet for 3 or 7 days and the effect on several hydroxylations involved in the biosynthesis and metabolism of bile acids was studied. The hydroxylations studied were all catalyzed by the microsomal fraction of liver homogenate fortified with NADPH. The 7α-hydroxylation of cholesterol was inhibited by feeding taurocholic acid, taurocheno-deoxycholic acid and taurodeoxycholic acid for 3 as well as 7 days. No marked inhibition was obtained with taurohyodeoxycholic acid or taurolithocholic acid. The 12α-hydroxylation of 7α-hydroxy-4-cholesten-3-one was inhibited after 3 as well as 7 days by all bile acids except taurohyodeoxycholic acid. With this acid a marked stimulation of 12α-hydroxylation was observed. The effects of the different bile acids on the 7α-hydroxylation of taurodeoxycholic acid were not very marked. The 6β-hydroxylation of lithocholie acid and taurochenodeoxycholic acid was stimulated by taurocholic acid and taurodeoxycholic acid. The reaction was inhibited by taurochenodeoxycholic acid, at least after 7 days. Taurohyodeoxycholic acid inhibited the 6β-hydroxylation slightly and taurolithocholic acid had no effect. The results were discussed in the light of present knowledge concerning mechanisms of regulation of formation and metabolism of bile acids and it was suggested that the mechanisms may be more complex than previously thought.     

Bile acid regulation of hepatic physiology: I. Hepatocyte transport of bile acids

Bile acids are cholesterol derivatives that serve as detergents in bile and the small intestine. Approximately 95% of bile acids secreted by hepatocytes into bile are absorbed from the distal ileum into the portal venous system. Extraction from the portal circulation by the hepatocyte followed by reexcretion into the bile canaliculus completes the enterohepatic circulation of these compounds. Over the past few years, candidate bile acid transport proteins of the sinusoidal and canalicular plasma membranes of the hepatocyte have been identified. The physiology of hepatocyte bile acid transport and its relationship to these transport proteins is the subject of this Themes article.     

Identification of unconjugated bile acids in human bile

Unconjugated bile acids in the bile of healthy and diseased humans were determined qualitatively and quantitatively by means of gas-liquid chromatography and gas-liquid chromatography-mass spectrometry, after their isolation by ion-exchange chromatography. In a healthy person and three patients with cholelithiasis, unconjugated bile acids comprised 0.1-0.4% of total biliary bile acids. The bile acid composition of the unconjugated fraction was quite different from that of the glycine- or taurine-conjugate fraction, in that it contained a relatively large proportion of unusual bile acids including C23 and C27 bile acids. In two patients with cerebrotendinous xanthomatosis, C22 and C23 bile acids were the major constituents of the biliary unconjugated bile acids, and comprised about 0.8% of total bile acids; no detectable amounts of C27 bile acids were found in their bile. The analysis of biliary unconjugated bile acids may be useful for the diagnosis of metabolic diseases concerning bile acids, particularly the accumulation or disappearance of unusual bile acids.     

Intracellular sterol transport and distribution

Sterols are important components of many biological membranes, and changes in sterol levels can have dramatic effects on membrane properties. Sterols are transported rapidly between cellular organelles by vesicular and nonvesicular processes. Recent studies have identified transmembrane proteins that facilitate the removal of sterols from membranes as well as soluble cytoplasmic proteins that play a role in their movement through the cytoplasm. The mechanisms by which these proteins work are generally not well understood. Cells maintain large differences in the sterol:phospholipid ratio in different organelles. Recent theoretical and experimental studies indicate ways in which the lipid environment can alter the chemical potential of sterols, which may help to explain aspects of their transport kinetics and distribution.     

Intracellular transport of human immunodeficiency virus

On entering a host cell, genomic components of human immunodeficiency virus (HIV) are translocated from plasma membrane to cell nucleus where the key events of the infectious process—virus genome integration into cell chromosomes and provirus formation—take place. After provirus expression, viral components move in the opposite direction, i.e., from nucleus to plasma membrane, for virus assembly. HIV translocation is provided by transport machinery of the host cell, which is strictly controlled by viral and cell proteins. Their functional activities are closely interrelated, while their interactions promote recognition and expression of translocation signals. The aim of this review is to consider functional capabilities of one of the main regulatory matrix proteins, MA. This virus-specific protein exhibits membranotropic and nucleophilic activities and controls intracellular movements of HIV throughout its life cycle. A hypothesis on the existence of two forms of MA and their functional roles is proposed. In-depth studies of intracellular targeting of HIV virions may shed additional light on intracellular transport pathways of HIV and identify new targets for anti-HIV drugs.     

Intracellular sorting and transport of proteins

The secretory and endocytic pathways of eukaryotic organelles consist of multiple compartments, each with a unique set of proteins and lipids. Specific transport mechanisms are required to direct molecules to defined locations and to ensure that the identity, and hence function, of individual compartments are maintained. The localisation of proteins to specific membranes is complex and involves multiple interactions. The recent dramatic advances in understanding the molecular mechanisms of membrane transport has been due to the application of a multi-disciplinary approach, integrating membrane biology, genetics, imaging, protein and lipid biochemistry and structural biology. The aim of this review is to summarise the general principles of protein sorting in the secretory and endocytic pathways and to highlight the dynamic nature of these processes. The molecular mechanisms involved in this transport along the secretory and endocytic pathways are discussed along with the signals responsible for targeting proteins to different intracellular locations.     

Intracellular transport and metabolism of sphingomyelin

SM is unique among the phospholipids because it is restricted to the lumenal aspect of organelles involved in the secretory and endocytic pathways. Given the intracellular sites of SM biosynthesis and hydrolysis, and the interconnections between these sites by vesicle-mediated transport pathways, the basic mechanism for maintaining the intracellular distribution of SM seems clear. It remains to be determined how SM metabolism and transport are coordinated to maintain the SM content of each organelle. For example, the size of the SM pool at the cell surface is maintained by regulation of at least five processes: transport of newly synthesized SM from the Golgi apparatus, plasma membrane lipid recycling, local SM synthesis, local SM hydrolysis, and SM transport from the cell surface to lysosomes. Although SM cannot undergo spontaneous transbilayer movement, SM metabolism generates both DAG, Cer and (indirectly) SPhB which can rapidly 'flip-flop', and thus gain access to the cytoplasmic leaflet of a membrane. It is of particular interest that these lipid species may be involved in the regulation of PK-C, suggesting that SM metabolism could play a role in signal transduction. However, physiological effects of endogenous Cer and SPhB remain elusive, even though the pharmacological effect of SPhB on PK-C is well established. Aside from the direct generation of second messengers, stimulation of SM hydrolysis has also been shown to induce cholesterol movement from the cell surface to intracellular membranes. It is not known whether this reflects the possibility that cholesterol may act as a second messenger. Alternatively, this phenomenon suggests that SM metabolism may cause rapid changes in the physical properties of the cell surface. For example, erythrocytes extensively treated with exogenously-added SMase will undergo endovesiculation It is tempting to speculate that any involvement of SM in the regulation of intracellular processes requires a combination of both the generation of biochemical second messengers and the alteration of membrane biophysical properties that can result from SM metabolism.