Biological functions of lysosomal membrane-associated glycoproteins

Lysosomes are the final repository of degradation products from the extracellular and intracellular spaces. The lysosomal membrane forms a unique vacuole participating in both endocytosis and autophagocytosis. It is extremely resistant to degradation by lysosomal hydrolases, maintains an acidic intralysosomal environment, transports amino acids and oligosaccharides produced by lysosomal hydrolases, interacts and fuses with other membrane organelles, such as endosomes and phagosomes, and with the plasma membrane.Biomedical Reviews 1997; 8: 119-125

SNAT7 is the primary lysosomal glutamine exporter required for extracellular protein-dependent growth of cancer cells

Significance Lysosomes are degradative intracellular organelles essential to cell maintenance and homeostasis. Although their degradative function is well documented, the proteins responsible for the efflux, and reuse, of lysosomal degradation products remain largely unknown. In this study, we identify the transporter responsible for lysosomal efflux of glutamine, an amino acid central to several key metabolic pathways. This central role of glutamine is exploited by several types of cancer cells with increased consumption of glutamine. Interestingly, genetic inactivation of the transporter impairs their growth under conditions of limited glutamine availability when internalized extracellular proteins are used as an alternative source of amino acids, suggesting novel approaches for anticancer therapies.</jats:p

A dileucine signal situated in the C-terminal tail of the lysosomal membrane protein p40 is responsible for its targeting to lysosomes

International audienceTransport of newly synthesised lysosomal membrane proteins from the TGN to the lysosomes is due to the presence of specific signals in their cytoplasmic domains that are recognised by cytosolic adaptors. p40, a hypothetical transporter of 372 amino acids localised in the lysosomal membrane, contains four putative lysosomal sorting motifs in its sequence: three of the YXXΦ–type (Y6QLF, Y106VAL, Y333NGL) and one of the [D/E]XXXL[L/I]-type (EQERL360L361). To test the implication of these motifs in the biosynthetic transport of p40, we replaced the most critical residues of these consensus sequences, the tyrosine or the leucine-leucine pair, by alanine or alanine-valine, respectively. We analysed the subcellular localisation of the mutated p40 proteins in transfected HeLa cells by confocal microscopy and by biochemical approaches (subcellular fractionation on self-forming Percoll density gradients and cell surface biotinylation). Our results show that p40 is mis-targeted to the plasma membrane when its dileucine motif is disrupted. No role of the tyrosine motifs could be put forward. Taken together, our results provide evidence that the sorting of p40 from the TGN to the lysosomes is directed by the dileucine EQERL360L361 motif situated in its C-terminal tail

Superconducting routing platform for large-scale integration of quantum technologies

To reach large-scale quantum computing, three-dimensional integration of scalable qubit arrays and their control electronics in multi-chip assemblies is promising. Within these assemblies, the use of superconducting interconnections, as routing layers, offers interesting perspective in terms of (1) thermal management to protect the qubits from control electronics self-heating, (2) passive device performance with significant increase of quality factors and (3) density rise of low and high frequency signals thanks to minimal dispersion. We report on the fabrication, using 200 mm silicon wafer technologies, of a multi-layer routing platform designed for the hybridization of spin qubit and control electronics chips. A routing level couples the qubits and the control circuits through one layer of Al0.995Cu0.005 and superconducting layers of TiN, Nb or NbN, connected between them by W-based vias. Wafer-level parametric tests at 300 K validate the yield of these technologies and low temperature electrical measurements in cryostat are used to extract the superconducting properties of the routing layers. Preliminary low temperature radio-frequency characterizations of superconducting passive elements, embedded in these routing levels, are presented

Loss of Niemann-Pick C1 or C2 Protein Results in Similar Biochemical Changes Suggesting That These Proteins Function in a Common Lysosomal Pathway

Niemann-Pick Type C (NPC) disease is a lysosomal storage disorder characterized by accumulation of unesterified cholesterol and other lipids in the endolysosomal system. NPC disease results from a defect in either of two distinct cholesterol-binding proteins: a transmembrane protein, NPC1, and a small soluble protein, NPC2. NPC1 and NPC2 are thought to function closely in the export of lysosomal cholesterol with both proteins binding cholesterol in vitro but they may have unrelated lysosomal roles. To investigate this possibility, we compared biochemical consequences of the loss of either protein. Analyses of lysosome-enriched subcellular fractions from brain and liver revealed similar decreases in buoyant densities of lysosomes from NPC1 or NPC2 deficient mice compared to controls. The subcellular distribution of both proteins was similar and paralleled a lysosomal marker. In liver, absence of either NPC1 or NPC2 resulted in similar alterations in the carbohydrate processing of the lysosomal protease, tripeptidyl peptidase I. These results highlight biochemical alterations in the lysosomal system of the NPC-mutant mice that appear secondary to lipid storage. In addition, the similarity in biochemical phenotypes resulting from either NPC1 or NPC2 deficiency supports models in which the function of these two proteins within lysosomes are linked closely

Early high antibody titre convalescent plasma for hospitalised COVID-19 patients: DAWn-plasma.

peer reviewedBACKGROUND: Several randomised clinical trials have studied convalescent plasma for coronavirus disease 2019 (COVID-19) using different protocols, with different severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralising antibody titres, at different time-points and severities of illness. METHODS: In the prospective multicentre DAWn-plasma trial, adult patients hospitalised with COVID-19 were randomised to 4 units of open-label convalescent plasma combined with standard of care (intervention group) or standard of care alone (control group). Plasma from donors with neutralising antibody titres (50% neutralisation titre (NT(50))) ≥1/320 was the product of choice for the study. RESULTS: Between 2 May 2020 and 26 January 2021, 320 patients were randomised to convalescent plasma and 163 patients to the control group according to a 2:1 allocation scheme. A median (interquartile range) volume of 884 (806-906) mL) convalescent plasma was administered and 80.68% of the units came from donors with neutralising antibody titres (NT(50)) ≥1/320. Median time from onset of symptoms to randomisation was 7 days. The proportion of patients alive and free of mechanical ventilation on day 15 was not different between both groups (convalescent plasma 83.74% (n=267) versus control 84.05% (n=137)) (OR 0.99, 95% CI 0.59-1.66; p=0.9772). The intervention did not change the natural course of antibody titres. The number of serious or severe adverse events was similar in both study arms and transfusion-related side-effects were reported in 19 out of 320 patients in the intervention group (5.94%). CONCLUSIONS: Transfusion of 4 units of convalescent plasma with high neutralising antibody titres early in hospitalised COVID-19 patients did not result in a significant improvement of clinical status or reduced mortality