Membrane interaction and disulphide-bridge formation in the unconventional secretion of Tau

Misfolded, pathological tau protein propagates from cell to cell causing neuronal degeneration in Alzheimer’s disease and other tauopathies. The molecular mechanisms of this process have remained elusive. Unconventional secretion of tau takes place via several different routes, including direct penetration through the plasma membrane. Here, we show that tau secretion requires membrane interaction via disulphide bridge formation. Mutating residues that reduce tau interaction with membranes or formation of disulphide bridges decrease both tau secretion from cells, and penetration through artificial lipid membranes. Our results demonstrate that tau is indeed able to penetrate protein-free membranes in a process independent of active cellular processes and that both membrane interaction and disulphide bridge formation are needed for this process. QUARK-based de novo modelling of the second and third microtubule-binding repeat domains (MTBDs), in which the two cysteine residues of 4R isoforms of tau are located, supports the concept that this region of tau could form transient amphipathic helices for membrane interaction.     

Cytoplasmic oxysterol-binding proteins: sterol sensors or transporters?

Families of oxysterol-binding protein (ORP) homologues are present in eukaryotes from yeast to man. Their hallmark feature is a characteristic ligand binding domain that, for several family members, has been shown to accommodate different oxysterols and/or cholesterol. ORPs of the “long” subtype contain targeting determinants for the endoplasmic reticulum and to other organelle membranes, the most prominent of which are phosphoinositide-binding pleckstrin homology domains, while “short” ORPs comprise a ligand binding domain with little additional sequences. There is increasing evidence that both long and short ORPs can be enriched at membrane contact sites, junctions of the endoplasmic reticulum with other organelles, where they are suggested to execute regulatory or sterol transfer functions. In this review we discuss the current evidence for putative roles of ORPs as sterol sensors or transporters.     

BODIPY-cholesterol: a new tool to visualize sterol trafficking in living cells and organisms

Analysis of sterol distribution and transport in living cells has been hampered by the lack of bright, photostable fluorescent sterol derivatives that closely resemble cholesterol. In this study, we employed atomistic simulations and experiments to characterize a cholesterol compound with fluorescent boron dipyrromethene difluoride linked to sterol carbon-24 (BODIPY-cholesterol). This probe packed in the membrane and behaved similarly to cholesterol both in normal and in cholesterol-storage disease cells and with trace amounts allowed the visualization of sterol movement in living systems. Upon injection into the yolk sac, BODIPY-cholesterol did not disturb zebrafish development and was targeted to sterol-enriched brain regions in live fish. We conclude that this new probe closely mimics the membrane partitioning and trafficking of cholesterol and, because of its excellent fluorescent properties, enables the direct monitoring of sterol movement by time-lapse imaging using trace amounts of the probe. This is, to our knowledge, the first cholesterol probe that fulfills these prerequisites.     

Allelopathic Effects of Toxic Haptophyte <Emphasis Type="Italic">Prymnesium parvum</Emphasis> Lead to Release of Dissolved Organic Carbon and Increase in Bacterial Biomass

The haptophyte Prymnesium parvum has lytic properties, and it affects coexisting phytoplankton species through allelopathy. We studied the effect of P. parvum allelochemicals on the lysis of the nontoxic and nonaxenic cryptomonad Rhodomonas salina and the consequent release of dissolved organic carbon (DOC). Changes in production, cell density, and biomass of associated bacteria were measured over 12 h. Six different combinations of P. parvum and R. salina cultures, their cell- and bacteria-free filtrates, and growth media as controls were used in the experiments. When P. parvum and R. salina cells were mixed, a significant increase in DOC concentration was measured within 30 min. Bacterial biomass increased significantly during the next 6 to 12 h when R. salina was mixed either with the P. parvum culture or the cell-free P. parvum filtrates (allelochemicals only). In contrast, bacterial biomass did not change in the treatments without the allelopathic action (without R. salina cells). Blooms of P. parvum alter the functioning of the planktonic food web by increasing carbon transfer through the microbial loop. In addition, P. parvum may indirectly benefit from the release of DOC as a result of its ability to ingest bacteria, by which it can acquire nutrients during limiting conditions.     

Rab8-dependent recycling promotes endosomal cholesterol removal in normal and sphingolipidosis cells

The mechanisms by which low-density lipoprotein (LDL)-cholesterol exits the endocytic circuits are not well understood. The process is defective in Niemann-Pick type C (NPC) disease in which cholesterol and sphingolipids accumulate in late endosomal compartments. This is accompanied by defective cholesterol esterification in the endoplasmic reticulum and impaired ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol efflux. We show here that overexpression of the recycling/exocytic Rab GTPase Rab8 rescued the late endosomal cholesterol deposition and sphingolipid mistrafficking in NPC fibroblasts. Rab8 redistributed cholesterol from late endosomes to the cell periphery and stimulated cholesterol efflux to the ABCA1-ligand apolipoprotein A-I (apoA-I) without increasing cholesterol esterification. Depletion of Rab8 from wild-type fibroblasts resulted in cholesterol deposition within late endosomal compartments. This cholesterol accumulation was accompanied by impaired clearance of LDL-cholesterol from endocytic circuits to apoA-I and could not be bypassed by liver X receptor activation. Our findings establish Rab8 as a key component of the regulatory machinery that leads to ABCA1-dependent removal of cholesterol from endocytic circuits.     

Neochloris oleoabundans oil production in an outdoor tubular photobioreactor at pilot scale

Journal of Applied Phycology - Oil production was tested with Neochloris oleoabundans in a 6&nbsp;m3, horizontal soft sleeve tubular reactor from 22 October to 7 November in Matalascañas,...     

Developing microalgal oil production for an outdoor photobioreactor

Journal of Applied Phycology - In this paper the preparations are described to develop a production of oil rich microalgal biomass under south European conditions. Ten microalgal species were...