Jürgen Pohl

A New Concept of Cellular Uptake and Intracellular Trafficking of Long-Chain Fatty Acids

Fatty acids are the main structural and energy sources of the human body. Within the organism, they are presented to cells as fatty acid: albumin complexes. Dissociation from albumin represents the first step of the cellular uptake process, involving membrane proteins with high affinity for fatty acids, e.g., fatty acid translocase (FAT/CD 36) or the membrane fatty acid-binding protein (FABPpm). According to the thus created transmembrane concentration gradient, uncharged fatty acids can flip-flop from the outer leaflet across the phospholipid bilayer. At the cytosolic surface of the plasma membrane, fatty acids can associate with the cytosolic FABP (FABPc) or with caveolin-1. Caveolins are constituents of caveolae, which are proposed to serve as lipid delivery vehicles for subcellular organelles. It is not known whether protein (FABPc)- and lipid (caveolae)-mediated intracellular trafficking of fatty acids operates in conjunction, or in parallel. Channeling fatty acids to the different metabolic pathways requires activation to acyl-CoA. For this process, the family of fatty acid transport proteins (FATP 1-5/6) might be relevant because they have been shown to possess acyl-CoA synthetase activity. Their variable N-terminal signaling sequences suggest that they might be targeted to specific organelles by anchoring in the phospholipid bilayer of the different subcellular membranes. At the highly conserved cytosolic AMP-binding site of FATP, fatty acids are activated to acyl-CoA for subsequent metabolic disposition by specific organelles. Overall, fatty acid uptake represents a continuous flow involving the following: dissociation from albumin by membrane proteins with high affinity for fatty acids; passive flip-flop across the phospholipid bilayer; binding to FABPc and caveolin-1 at the cytosolic plasma membrane; and intracellular trafficking via FABPc and/or caveolae to sites of metabolic disposition. The uptake process is terminated after activation to acyl-CoA by the members of the FATP family targeted intracellularly to different organelles.

Translocation of long chain fatty acids across the plasma membrane - lipid rafts and fatty acid transport proteins

Translocation of long chain fatty acids across the plasma membrane is achieved by a concert of co-existing mechanisms. These lipids can passively diffuse, but transport can also be accelerated by certain membrane proteins as well as lipid rafts. Lipid rafts are dynamic assemblies of proteins and lipids, that float freely within the two dimensional matrix of the membrane bilayer. They are receiving increasing attention as devices that regulate membrane function in vivo and play an important role in membrane trafficking and signal transduction. In this review we will discuss how lipid rafts might be involved in the uptake process and how the candidate proteins for fatty acid uptake FAT/CD36 and the FATP proteins interact with these domains. We will also discuss the functional role of FATPs in general. To our understanding FATPs are indirectly involved in the translocation process across the plasma membrane by providing long chain fatty acid synthetase activity.

New concepts of cellular fatty acid uptake: role of fatty acid transport proteins and of caveolae

Efficient uptake and channelling of long-chain fatty acids (LCFA) are critical cell functions. Evidence is emerging that proteins are important mediators of LCFA-trafficking into cells and various proteins have been suggested to be involved in this process. Amongst these proteins is a family of membrane-associated proteins termed fatty acid transport proteins (FATP). So far six members of this family, designated FATP 1-6, have been characterized. FATP 1, 2 and 6 show a highly-conserved AMP-binding region that participates in the activation of very-long-chain fatty acids (VLCFA) to form their acyl-CoA derivatives. The mechanisms by which FATP mediate LCFA uptake are not well understood, but several studies provide evidence that uptake of LCFA across cellular membranes is closely linked to acyl-CoA synthetase activity. It is proposed that FATP indirectly enhance LCFA uptake by activating VLCFA to their CoA esters, which are required to maintain the typical structure of lipid rafts in cellular membranes. Recent work has shown that the structural integrity of lipid rafts is essential for cellular LCFA uptake. This effect might be exerted by proteins, e.g. caveolin-1 and FAT/CD36, that use lipid rafts as platforms and bind or transport LCFA. The proposed molecular mechanisms await further experimental investigation.

Group B streptococcal β-hemolysin induces mortality and liver injury in experimental sepsis

New Zealand White rabbits were challenged with the wild-type (wt) group B streptococci (GBS) serotype III strain (COH1) and its isogenic nonhemolytic (NH) and hyperhemolytic (HH) mutants. Mortality differed significantly between rabbits infected with the HH mutant IN40 (67%), compared with rabbits infected with the wt COH1 strain (27%) and the NH strains COH1-20 and COH1:cylEDeltacat (13% and 0%, respectively; P<.05). Histopathologically, disseminated septic microabscesses surrounded by necrotic foci were found exclusively in the livers of HH mutant IN40-infected animals. Serum transaminase levels were 20-fold higher in the HH-infected group, compared with rabbits infected with the other strains. Positive TUNEL (in situ terminal deoxynucleotide transferase-mediated dUTP nick end labeling) staining and activation of caspase-3 in hepatocytes were more frequent in HH-infected than in wt-infected animals and absent in the NH mutant COH1-20-infected group, indicating that GBS beta-hemolysin triggers apoptotic pathways in hepatocytes. This work provides the first evidence that GBS beta-hemolysin plays a crucial role in the pathophysiology of GBS sepsis by inducing liver failure and high mortality.

Systemic Chemotherapy with Epirubicin for Treatment of Advanced or Multifocal Hepatocellular Carcinoma

Background: The purpose of this retrospective study was to determine the response rate and effect on survival of chemotherapy with epirubicin in non-resectable advanced hepatocellular carcinoma (HCC). Methods: Fifty-two patients with non-resectable disease were treated with epirubicin. A treatment cycle consisted of 20 mg/m2 i.v. on days 1, 8 and 15 and was repeated every 4 weeks to a maximum dose of 1,000 mg/m2. Forty-four patients were eligible for analysis. Results: Out of 44 patients, 1 (2.3%) achieved a complete response, 3 (6.8%) had partial responses and 16 (36%) had stable disease (SD). For patients with successful disease control (complete and partial responders and patients with SD), the median survival was 16.2 months; for non-responders, it was 6.1 months (p < 0.003). Eight (88.9%) of 9 patients with alpha-fetoprotein (AFP) levels <50 µg/l achieved successful disease control compared to 12 (34.9%) out of 35 patients with initially elevated AFP (p < 0.0001). Conclusion: Epirubicin appears to be an active therapeutic option for patients with non-resectable HCC. Especially the subgroup of patients with low levels of AFP may benefit from this treatment.

Role of FATP in parenchymal cell fatty acid uptake

Long-chain fatty acids (LCFAs) represent key metabolites for energy generation and storage. Transport and metabolism of LCFA are believed to be regulated by membrane-associated proteins that bind and transport LCFA. Identifying the postulated fatty acid transporters is of considerable interest since altered fatty acid uptake has been implicated in disease such as insulin resistance and obesity. Recently, a family of membrane associated proteins, termed fatty acid transport proteins (FATPs), have been described that enhance uptake of LCFAs. Until today, six members of this family, designated FATP1-6, have been characterized. This review will focus on FATP structure, expression patterns, regulation, mechanism of transport and clinical implications.