Does the Food Ingredient Pectin Provide a Risk for Patients Allergic to Non-Specific Lipid-Transfer Proteins?

Pectin, a dietary fiber, is a polysaccharide that is widely used in food industry as a gelling agent. In addition, prebiotic and beneficial immunomodulatory effects of pectin have been demonstrated, leading to increased importance as food supplement. However, as cases of anaphylactic reactions after consumption of pectin-supplemented foods have been reported, the present study aims to evaluate the allergy risk of pectin. This is of particular importance since most of the pectin used in the food industry is extracted from citrus or apple pomace. Both contain several allergens such as non-specific lipid transfer proteins (nsLTPs), known to induce severe allergic reactions, which could impair the use of pectins in nsLTP allergic patients. Therefore, the present study for the first time was performed to analyze residual nsLTP content in two commercial pectins using different detection methods. Results showed the analytical sensitivity was diminished by the pectin structure. Finally, spiking of pectin with allergenic peach nsLTP Pru p 3 led to the conclusion that the potential residual allergen content in both pectins is below the threshold to induce anaphylactic reactions in nsLTP-allergic patients. This data suggests that consumption of the investigated commercial pectin products provides no risk for inducing severe reactions in nsLTP-allergic patients.

Functional analyses of phosphatidylserine/PI(4)P exchangers with diverse lipid species and membrane contexts reveal unanticipated rules on lipid transfer

Background Lipid species are accurately distributed in the eukaryotic cell so that organelle and plasma membranes have an adequate lipid composition to support numerous cellular functions. In the plasma membrane, a precise regulation of the level of lipids such as phosphatidylserine, PI(4)P, and PI(4,5)P2, is critical for maintaining the signaling competence of the cell. Several lipid transfer proteins of the ORP/Osh family contribute to this fine-tuning by delivering PS, synthesized in the endoplasmic reticulum, to the plasma membrane in exchange for PI(4)P. To get insights into the role of these PS/PI(4)P exchangers in regulating plasma membrane features, we question how they selectively recognize and transfer lipid ligands with different acyl chains, whether these proteins exchange PS exclusively for PI(4)P or additionally for PI(4,5)P2, and how sterol abundance in the plasma membrane impacts their activity. Results We measured in vitro how the yeast Osh6p and human ORP8 transported PS and PI(4)P subspecies of diverse length and unsaturation degree between membranes by fluorescence-based assays. We established that the exchange activity of Osh6p and ORP8 strongly depends on whether these ligands are saturated or not, and is high with representative cellular PS and PI(4)P subspecies. Unexpectedly, we found that the speed at which these proteins individually transfer lipid ligands between membranes is inversely related to their affinity for them and that high-affinity ligands must be exchanged to be transferred more rapidly. Next we determined that Osh6p and ORP8 cannot use PI(4,5)P2 for exchange processes, because it is a low-affinity ligand, and do not transfer more PS into sterol-rich membranes. Conclusions Our study provides new insights into PS/PI(4)P exchangers by indicating the degree to which they can regulate the acyl chain composition of the PM, and how they control PM phosphoinositide levels. Moreover, we establish general rules on how the activity of lipid transfer proteins relates to their affinity for ligands.

ORP5 AND ORP8 ORCHESTRATE LIPID DROPLET BIOGENESIS AND MAINTENANCE AT ER-MITOCHONDRIA CONTACT SITES

Lipid droplets (LDs) are the primary organelles of lipid storage, buffering energy fluctuations of the cell. They store neutral lipids in their core that is surrounded by a protein-decorated phospholipid monolayer. LDs arise from the Endoplasmic Reticulum (ER). The ER-protein seipin, localizing at ER-LD junctions, controls LD nucleation and growth. However, how LD biogenesis is spatially and temporally coordinated remains elusive. Here, we show that the lipid transfer proteins ORP5 and ORP8 control LD biogenesis at Mitochondria-Associated ER Membrane (MAM) subdomains, enriched in phosphatidic acid. We found that ORP5/8 regulate seipin recruitment to these MAM-LD contacts, and their loss impairs LD biogenesis. Importantly, the integrity of ER-mitochondria contact sites is crucial for the ORP5/8 function in regulating seipin-mediated LD biogenesis. Our study uncovers an unprecedented ORP5/8 role in orchestrating LD biogenesis at MAMs and brings novel insights into the metabolic crosstalk between mitochondria, ER, and LDs at membrane contact sites.

Transcriptomic Responses of Potato to Drought Stress

Transcriptomic changes in plants during drought stress give insights into the mechanisms with which plants stabilise their metabolic processes in order to cope with the drought condition. In potato, understanding such drought-induced transcriptomic changes is critical because prolonged field drought interferes with tuber formation and bulking period of potato development, which eventually affects yield. We hypothesised that phenotypic drought responses of potato genotypes may be linked to differences in transcriptomic changes. Using an RNA sequencing approach, we investigated such transcriptomic changes in leaves of three cultivars (Biogold, Hansa and Lady Rosetta) under drought. We found more differentially expressed genes (DEGs) in the tolerant cultivars, Lady Rosetta and Biogold, than in the sensitive cultivar (Hansa). The differential gene expression trend reflected the phenotypic drought responses of the cultivars. For instance, we found in both Biogold and Lady Rosetta but not in Hansa, an upregulation of genes involved in carbohydrate metabolism (e.g., Alpha-glucosidase), flavonoid biosynthesis (e.g., Flavanone 3 beta-hydroxylase), lipid biosynthesis/transfer (e.g., nonspecific Lipid Transfer Proteins), heat shock proteins and secondary metabolites like phenolics and lignins. Furthermore, a prolonged drought stress resulted in reduced DEGs in Biogold and Hansa, but not in Lady Rosetta that also maintained its tuber yield under such prolonged stress suggesting a more robust drought tolerance. Our findings suggest that a synergistic expression of genes involved in several different aspects of drought response is required in order to obtain a robust tolerance.

Transport Pathways That Contribute to the Cellular Distribution of Phosphatidylserine

Phosphatidylserine (PS) is a negatively charged phospholipid that displays a highly uneven distribution within cellular membranes, essential for establishment of cell polarity and other processes. In this review, we discuss how combined action of PS biosynthesis enzymes in the endoplasmic reticulum (ER), lipid transfer proteins (LTPs) acting within membrane contact sites (MCS) between the ER and other compartments, and lipid flippases and scramblases that mediate PS flip-flop between membrane leaflets controls the cellular distribution of PS. Enrichment of PS in specific compartments, in particular in the cytosolic leaflet of the plasma membrane (PM), requires input of energy, which can be supplied in the form of ATP or by phosphoinositides. Conversely, coupling between PS synthesis or degradation, PS flip-flop and PS transfer may enable PS transfer by passive flow. Such scenario is best documented by recent work on the formation of autophagosomes. The existence of lateral PS nanodomains, which is well-documented in the case of the PM and postulated for other compartments, can change the steepness or direction of PS gradients between compartments. Improvements in cellular imaging of lipids and membranes, lipidomic analysis of complex cellular samples, reconstitution of cellular lipid transport reactions and high-resolution structural data have greatly increased our understanding of cellular PS homeostasis. Our review also highlights how budding yeast has been instrumental for our understanding of the organization and transport of PS in cells.

A tetrameric assembly of saposin A: increasing structural diversity in lipid transfer proteins

Saposins are lipid transfer proteins required for the degradation of sphingolipids in the lysosome. These small proteins bind lipids by transitioning from a closed, monomeric state to an open conformation exposing a hydrophobic surface that binds and shields hydrophobic lipid tails from the aqueous environment. Saposins form a range of multimeric assemblies to encompass these bound lipids and present them to hydrolases in the lysosome. This lipid-binding property of human saposin A has been exploited to form lipoprotein nanodiscs suitable for structural studies of membrane proteins. Here we present the crystal structure of a unique tetrameric assembly of murine saposin A produced serendipitously, following modifications of published protocols for making lipoprotein nanodiscs. The structure of this new saposin oligomer highlights the diversity of tertiary arrangement that can be adopted by these important lipid transfer proteins.

Anaphylaxis to Sunflower Seed with Tolerance to Sunflower Oil: A Case Report

Sunflower seeds (Helianthus annuus) are an uncommon source of allergy; however, some cases of allergy to sunflower seeds have been reported. Sunflower seed sensitization occurs to storage proteins (2S albumins) and lipid transfer proteins (LTPs). A 46-year-old female presented three allergic reactions within minutes of consuming sunflower seeds. A prick-to-prick test indicated a positive reaction only to sunflower seeds and a negative reaction to other nuts, such as almond, hazelnut, pistachio, cashew, peanut, macadamia, sesame, and walnut. Prick-to-prick and oral provocation tests of sunflower oil were performed, and a negative result was obtained. The patient was prescribed a 0.3 mg epinephrine autoinjector device for emergency intramuscular administration. The patient is currently under avoidance of sunflower seed but eats food cooked in sunflower seed oil. Based on this case, we should recognize that sunflower seeds have the potential to cause severe anaphylaxis, which indicates tolerance to sunflower oil. An accurate and fast diagnosis allows timely recommendation to practice strict avoidance of sunflower seeds, thus reducing the possibility of recurrence of an anaphylactic reaction.