Stabilization of sphingomyelin interactions by interfacial hydroxyls — A study of phytosphingomyelin properties

AbstractD-ribo-phytosphingosines are biologically significant long-chain bases present in various sphingolipids from yeasts, fungi, plants and mammals. In this study we prepared phytopalmitoylsphingomyelin (phytoPSM) analogs based on the D-ribo-phytosphingosine base. The N-linked acyl chains were either 16:0, 2OH(R)16:0 (natural isomer), or 2OH(S)16:0. The gel-phase of phytoPSM was more stable than that of PSM (Tm 48.6°C and 41.0°C, respectively). The gel-liquid crystalline phase transition enthalpies were 9.1±0.4 and 6.1±0.3kcal/mol for phytoPSM and PSM, respectively. An N-linked 2OH(R)16:0 in phytoPSM destabilized the gel phase relative to phytoPSM (by ~+6°C, based on DPH anisotropy measurements), whereas 2OH(S)16:0 in phytoPSM stabilized it (by ~−6°C). All phytoPSM analogs formed sterol-enriched ordered domains in a fluid ternary bilayer, and those containing phytoPSM or 2OH(S)phytoPSM were more thermostable than the domains containing 2OH(R)phytoPSM or PSM. The affinity of cholestatrienol for POPC bilayers containing 20mol% phytoPSM was higher than for comparable bilayers with an equal amount of PSM. The 2-hydroxylated acyl chains in phytoPSM did not markedly alter sterol affinity. We conclude that phytoPSM is a more ordered sphingolipid than PSM, and is fully capable of interacting with cholesterol

Membrane Properties of D-erythro-N-acyl Sphingomyelins and Their Corresponding Dihydro Species

AbstractWe have prepared acyl chain-defined D-erythro-sphingomyelins and D-erythro-dihydrosphingomyelins and compared their properties in monolayer and bilayer membranes. Surface pressure/molecular area isotherms of D-erythro-N-16:0-sphingomyelin (16:0-SM) and D-erythro-N-16:0-dihydrosphingomyelin (16:0-DHSM) show very similar packing properties, except that the expanded-to-condensed phase transition (crystallization) occurs at a lower surface pressure for 16:0-DHSM. The measured surface potential was generally about 100mV less for 16:0-DHSM monolayers compared to 16:0-SM monolayers. The condensed domains (crystals) that formed in 16:0-SM monolayers as a function of compression displayed star-shaped morphology when viewed under an epifluorescence microscope. 16:0-DHSM monolayers did not form similar crystals upon compression. 16:0-DHSM was degraded much faster by sphingomyelinase from Staphylococcus aureus than 16:0-SM (10-fold difference in enzyme activity needed for comparable hydrolytic rate). Cholesterol desorption from 16:0-DHSM to cyclodextrin was slightly slower (∼20%) than the rate measured from 16:0-SM monolayers (at 60mol % cholesterol). The bilayer melting temperature of 16:0-DHSM was 47.7°C (ΔH 8.3kcal/mol) whereas it was 41.2°C for 16:0-SM (ΔH 8.1kcal/mol). Cholesterol/16:0-DHSM bilayers (15mol % sterol) had more condensed domains than comparable 16:0-SM bilayers, as evidenced from the quenching resistance of DPH in DHSM membranes. We conclude that cholesterol interacts more favorably with 16:0-DHSM and that the membranes are more condensed than comparable 16:0-SM-containing membranes

Fine-scale observations of physical and biological environment along a herring feeding migration route

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Melle, W., Klevjer, T., Strand, E., Wiebe, P. H., Slotte, A., & Huse, G. Fine-scale observations of physical and biological environment along a herring feeding migration route. Deep-Sea Research Part II: Topical Studies in Oceanography, 180, (2020): 104845, doi:10.1016/j.dsr2.2020.104845.We observed herring horizontal and vertical distribution during feeding migration along a 128 km transect across the Arctic front of the Norwegian and Iceland seas, in early June, in relation to its physical, chemical and biological environment, distribution of prey organisms and pelagic and mesopelagic competitors. The Norwegian Spring Spawning herring is one of the largest and economically most important stocks of pelagic fish in the world and understanding what controls its feeding migration is, and has been for centuries, a major research question that also has major implications for management. High resolution ecosystem data were obtained by hull mounted multi-frequency acoustics and a towed platform undulating between 10 and 400 m equipped with multi-frequency acoustics, temperature, salinity and fluorescence sensors, an Optical Plankton Counter and a Video Plankton Recorder. Additional sampling was done by MOCNESS, Macroplankton trawl, and CTD equipped with water bottles for temperature, salinity, nutrients and chlorophyll at discrete stations along the transect. Biological characteristics and stomach content of the herring were obtained from samples at discrete trawl stations. The Arctic front proved to be an important transitional zone in zooplankton biomass, abundance and diversity. Phenology of phyto- and zooplankton also changed across the front, being somewhat delayed on the cold side. The herring were distributed all along the transect showing a shallow distribution on the warm side and both deep and shallow on the cold side, not clearly related to light and time of the day. The herring stomach content was higher on the cold side. There was no significant pattern in average age, weight, or body length of the herring along the transect. The herring were present and fed in the area of the transect during the time when the overwintering generation of Calanus finmarchicus dominated, before the development of the new generation of the year. We suggest that the phenology of C. finmarchicus can be an important driver of the herring feeding migration. While prey-availability was higher on the Arctic side of the front, light conditions for visual feeding at depth were probably better on the Atlantic side. The herring did not show classical dial vertical migration, but its prey did, and the herring's prey were probably available within the upper 100 m during the course of a 24 h cycle. With a general westward direction of migration, the herring along the transect moved towards lower temperatures and temperature did not seem to be a probable driver for migration. We conclude that fine-scale studies of herring migration and feeding can increase our understanding of the migratory processes and add to our understanding of large-scale distributional patterns, changes therein, and herring trophodynamics and ecological role. The fine-resolution parameters can also be important as input to ecosystem models.We would also like to acknowledge the funding from Euro-BASIN, EU FP7, Grant agreement No 264933, HARMES, Research Council of Norway project number 280546 and MEESO, EU H2020 research and innovation programme, Grant Agreement No 817669

An autocrine sphingosine-1-phosphate signaling loop enhances NF-kappaB-activation and survival

Peer reviewe

The Effect of Cholesterol on the Long-Range Network of Interactions Established among Sea Anemone Sticholysin II Residues at the Water-Membrane Interface

Actinoporins are α-pore forming proteins with therapeutic potential, produced by sea anemones. Sticholysin II (StnII) from Stichodactyla helianthus is one of its most extensively characterized members. These proteins remain stably folded in water, but upon interaction with lipid bilayers, they oligomerize to form a pore. This event is triggered by the presence of sphingomyelin (SM), but cholesterol (Chol) facilitates pore formation. Membrane attachment and pore formation require changes involving long-distance rearrangements of residues located at the protein-membrane interface. The influence of Chol on membrane recognition, oligomerization, and/or pore formation is now studied using StnII variants, which are characterized in terms of their ability to interact with model membranes in the presence or absence of Chol. The results obtained frame Chol not only as an important partner for SM for functional membrane recognition but also as a molecule which significantly reduces the structural requirements for the mentioned conformational rearrangements to occur. However, given that the DOPC:SM:Chol vesicles employed display phase coexistence and have domain boundaries, the observed effects could be also due to the presence of these different phases on the membrane. In addition, it is also shown that the Arg51 guanidinium group is strictly required for membrane recognition, independently of the presence of Chol

Toxin-induced pore formation is hindered by intermolecular hydrogen bonding in sphingomyelin bilayers

Sticholysin I and II (StnI and StnII) are pore-forming toxins that use sphingomyelin (SM) for membrane binding. We examined how hydrogen bonding among membrane SMs affected the StnI- and StnII-induced pore formation process, resulting in bilayer permeabilization. We compared toxin-induced permeabilization in bilayers containing either SM or dihydro-SM (lacking the trans 4 double bond of the long-chain base), since their hydrogen-bonding properties are known to differ greatly. We observed that whereas both StnI and StnII formed pores in unilamellar vesicles containing palmitoyl-SM or oleoyl-SM, the toxins failed to similarly form pores in vesicles prepared from dihydro-PSM or dihydro-OSM. In supported bilayers containing OSM, StnII bound efficiently, as determined by surface plasmon resonance. However, StnII binding to supported bilayers prepared from dihydro-OSM was very low under similar experimental conditions. The association of the positively charged StnII (at pH 7.0) with unilamellar vesicles prepared from OSM led to a concentration-dependent increase in vesicle charge, as determined from zeta-potential measurements. With dihydro-OSM vesicles, a similar response was not observed. Benzyl alcohol, which is a small hydrogen-bonding compound with affinity to lipid bilayer interfaces, strongly facilitated StnII-induced pore formation in dihydro-OSM bilayers, suggesting that hydrogen bonding in the interfacial region originally prevented StnII from membrane binding and pore formation. We conclude that interfacial hydrogen bonding was able to affect the membrane association of StnI- and StnII, and hence their pore forming capacity. Our results suggest that other types of protein interactions in bilayers may also be affected by hydrogen-bonding origination from SMs

Sticholysin, Sphingomyelin, and Cholesterol: A Closer Look at a Tripartite Interaction

Actinoporins are a group of soluble toxic proteins that bind to membranes containing sphingomyelin (SM) and oligomerize to form pores. Sticholysin II (StnII) is a member of the actinoporin family, produced by Stichodactyla helianthus. Cholesterol (Chol) is known to enhance the activity of StnII. However, the molecular mechanisms behind this activation have remained obscure, although the activation is not Chol specific but rather sterol specific. To further explore how bilayer lipids affect or are affected by StnII, we have used a multiprobe approach (fluorescent analogs of both Chol and SM) in combination with a series of StnII tryptophan (Trp)-mutants, to study StnII/bilayer interactions. First we compared StnII bilayer permeabilization in the presence of Chol or oleoyl-ceramide (OCer). The comparison was done since both Chol and OCer have a 1-hydroxyl which help to orient the molecule in the bilayer (although OCer have additional polar functional groups). Both Chol and OCer also have increased affinity for SM, which StnII may recognize. However, our results show that only Chol was able to activate StnII-induced bilayer permeabilization – OCer failed to active. To further examine possible Chol/StnII interactions, we measured Förster resonance energy transfer (FRET) between Trp in StnII and cholestatrienol (CTL), a fluorescent analog of Chol. We could show higher FRET efficiency between CTL and Trp:s in position 100 and 114 of StnII, when compared to three other Trp positions further away from the bilayer binding region of StnII. Taken together, our results suggest that StnII was able to attract Chol to its vicinity, maybe by showing affinity for Chol. SM interactions are known to be important for StnII binding to bilayers, and Chol is known to facilitate subsequent permeabilization of the bilayers by StnII. Our results help to better understand the role of these important membrane lipids for the bilayer properties of StnII

Structure and functioning of four North Atlantic ecosystems - A comparative study

The epi- and mesopelagic ecosystems of four sub-polar ocean basins, the Labrador, Irminger, Iceland and Norwegian seas, were surveyed during two legs from Bergen, Norway, to Nuuk, Greenland, and back to Bergen. The survey was conducted from 1 May to 14 June, and major results were published in five papers (Drinkwater et al., Naustvoll et al., Strand et al., Melle et al., this issue, and Klevjer et al., this issue a, this issue b). In the present paper, the structures of the ecosystem are reviewed, and aspects of the functioning of the ecosystems examined, focusing on a comparison of trophic relationships in the four basins. In many ways, the ecosystems are similar, which is not surprising since they are located at similar latitudes and share many hydrographic characteristics, like input of both warm and saline Atlantic water, as well as cold and less saline Arctic water. Literature review suggests that total annual primary production is intermediate in the eastern basins and peaks in the Labrador Sea, while the Irminger Sea is the most oligotrophic sea. This was not reflected in the measurements of different trophic levels taken during the cruise. The potential new production was estimated to be higher in the Irminger Sea than in the eastern basins, and while the biomass of mesozooplankton was similar across basins, the biomass of mesopelagic micronekton was about one order of magnitude higher in the western basins, and peaked in the Irminger Sea, where literature suggests annual primary production is at its lowest. The eastern basins hold huge stocks of pelagic planktivore fish stocks like herring, mackerel and blue whiting, none of which are abundant in the western seas. As both epipelagic nekton and mesopelagic micronekton primarily feed on the mesozooplankton, there is likely competitive interactions between the epipelagic and mesopelagic, but we're currently unable to explain the estimated ~1 order of magnitude difference in micronekton standing stock. The results obtained during the survey highlight that even if some aspects of pelagic ecosystems are well understood, we currently do not understand overall pelagic energy flow in the North Atlantic.publishedVersio