Single-molecule kinetics of pore assembly by the membrane attack complex

The membrane attack complex (MAC) is a hetero-oligomeric protein assembly that kills pathogens by perforating their cell envelopes. The MAC is formed by sequential assembly of soluble complement proteins C5b, C6, C7, C8 and C9, but little is known about the rate-limiting steps in this process. Here, we use rapid atomic force microscopy (AFM) imaging to show that MAC proteins oligomerize within the membrane, unlike structurally homologous bacterial pore-forming toxins. C5b-7 interacts with the lipid bilayer prior to recruiting C8. We discover that incorporation of the first C9 is the kinetic bottleneck of MAC formation, after which rapid C9 oligomerization completes the pore. This defines the kinetic basis for MAC assembly and provides insight into how human cells are protected from bystander damage by the cell surface receptor CD59, which is offered a maximum temporal window to halt the assembly at the point of C9 insertion

Emerging Molecular Mechanisms of Signal Transduction in Pentameric Ligand-Gated Ion Channels

International audienceNicotinic acetylcholine, serotonin type 3, γ-amminobutyric acid type A, and glycine receptors are major players of human neuronal communication. They belong to the family of pentameric ligand-gated ion channels, sharing a highly conserved modular 3D structure. Recently, high-resolution structures of both open- and closed-pore conformations have been solved for a bacterial, an invertebrate, and a vertebrate receptor in this family. These data suggest that a common gating mechanism occurs, coupling neurotransmitter binding to pore opening, but they also pinpoint significant differences among subtypes. In this Review, we summarize the structural and functional data in light of these gating models and speculate about their mechanistic consequences on ion permeation, pathological mutations, as well as functional regulation by orthosteric and allosteric effectors

Photoelectrochemical alcohols oxidation over polymeric carbon nitride photoanodes with simultaneous H2 production

The photoelectrochemical oxidation of organic molecules into valuable chemicals is a promising technology, but its development is hampered by the poor stability of photoanodic materials in aqueous solutions, low faradaic efficiency, low product selectivity, and a narrow working pH range. Here, we demonstrate the synthesis of value-added aldehydes and carboxylic acids with clean hydrogen (H) production in water using a photoelectrochemical cell based solely on polymeric carbon nitride (CN) as the photoanode. Isotope labeling measurements and DFT calculations reveal a preferential adsorption of benzyl alcohol and molecular oxygen to the CN layer, enabling fast proton abstraction and oxygen reduction, which leads to the synthesis of an aldehyde at the first step. Further oxidation affords the corresponding acid. The CN photoanode exhibits excellent stability (>40 h) and activity for the oxidation of a wide range of substituted benzyl alcohols with high yield, selectivity (up to 99%), and faradaic efficiency (>90%).This project has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. [849068]). This work was also partially supported by the joint Israel Science Foundation–National Science Foundation of China (ISF–NSFC), grant No. 2969/19, the ISF grant No.601/21, and by the Pazy Foundation (grant No. 119-2020). J.A. and H.G. gratefully acknowledge financial support by the Spanish Ministry of Science and Innovation (RTI2018-98237-CO2-1) and Generalitat Valenciana (Prometeo 2021/038). The authors thank Liel Abisdris, Jonathan Tzadikov, Ayelet Tashakory, and Rotem Geva for help with material characterization. We thank Dr Chabanne for fruitful discussion

An L,L- diaminopimelate aminotransferase mutation leads to metabolic shifts and growth inhibition in Arabidopsis

Lysine (Lys) connects the mitochondrial electron transport chain to amino acid catabolism and the tricarboxylic acid cycle. However, our understanding of how a deficiency in Lys biosynthesis impacts plant metabolism and growth remains limited. Here, we used a previously characterized Arabidopsis mutant (dapat) with reduced activity of the Lys biosynthesis enzyme L,L-diaminopimelate aminotransferase to investigate the physiological and metabolic impacts of impaired Lys biosynthesis. Despite displaying similar stomatal conductance and internal CO2 concentration, we observed reduced photosynthesis and growth in the dapat mutant. Surprisingly, whilst we did not find differences in dark respiration between genotypes, a lower storage and consumption of starch and sugars was observed in dapat plants. We found higher protein turnover but no differences in total amino acids during a diurnal cycle in dapat plants. Transcriptional and two-dimensional (isoelectric focalization/SDS-PAGE) proteome analyses revealed alterations in the abundance of several transcripts and proteins associated with photosynthesis and photorespiration coupled with a high glycine/serine ratio and increased levels of stress-responsive amino acids. Taken together, our findings demonstrate that biochemical alterations rather than stomatal limitations are responsible for the decreased photosynthesis and growth of the dapat mutant, which we hypothesize mimics stress conditions associated with impairments in the Lys biosynthesis pathway

Structure and evolution of an active resurgent dome evidenced by geophysical investigations: The Yenkahe dome-Yasur volcano system (Siwi caldera, Vanuatu)

Brothelande, E. et. al.In this contribution, we focus on one of the most active resurgences on Earth, that of the Yenkahe dome in the Siwi caldera (Tanna Island, Vanuatu), which is associated with the persistently active Yasur volcano. Gravity and magnetic surveys have been carried out over the past few years in the area, as well as electrical methods including electrical resistivity tomography (ERT), time domain electro-magnetics (TDEM) and self-potential (SP). These investigations were completed by thermometry, CO2 soil gas measurements, field observations and sampling. This multi-method approach allows geological structures within the caldera to be identified, as well as associated hydrothermal features. The global structure of the caldera is deduced from gravity data, which shows the caldera rim as a high density structure. Large lava fields, emplaced before and after the onset of resurgence, are evidenced by combined gravity, magnetic and resistivity signals. In the middle of the caldera, the Yenkahe dome apparently results from a combination of volcanic and tectonic events, showing that lava extrusion and resurgence have been operating simultaneously or alternately during the Siwi caldera post-collapse history. There is a clear distinction between the western and eastern parts of the dome. The western part is older and records the growth of an initial volcanic cone and the formation of a small caldera. This small caldera (paleo-Yasur caldera), partially filled with lava flows, is the present-day focus of volcanic activity and associated fluid circulation and alteration. The eastern part of the dome is presumably younger, and is characterized by intense, extensive hydrothermal alteration and activity. Its northern part is covered by lava flow piles and exhibits a shallow hydrothermal zone in ERT. The southern part has hydrothermal alteration and activity extending at least down to the base of the resurgent dome. This part of the dome is built up of low cohesion rock and is thus potentially prone to gravitational landslides. Lastly, while self-potential and temperature data suggest that widespread hydrothermal circulation occurs throughout almost all of the caldera, and possibly beyond, the most active parts of this hydrothermal system are associated with the dome. The presence of this active hydrothermal system is the clearest indicator that these methods can provide of a potential shallow magmatic body underneath the domeThis research was co-financed by the French Government “ANR ARC-Vanuatu: Programme Catastrophes Telluriques et Tsunamis” (ANR-06-CATT-02) and Laboratory of Excellence initiative n°ANR-10-LABX-0006, the Laboratoire Magmas et Volcans (LMV), the Laboratoire GéoSciences Réunion, the Région Auvergne and the European Regional Development Fund. Datasets are available at LMV. In addition, gravimetric data are available at International Gravimetric Bureau. This is Laboratory of Excellence ClerVolc contribution number 171 and IPGP contribution number 3654.Peer reviewe

Structure and evolution of an active resurgent dome evidenced by geophysical investigations: The Yenkahe dome-Yasur volcano system (Siwi caldera, Vanuatu)

International audienceIn this contribution, we focus on one of the most active resurgences on Earth, that of the Yenkahe dome in the Siwi caldera (Tanna Island, Vanuatu), which is associated with the persistently active Yasur volcano. Gravity and magnetic surveys have been carried out over the past few years in the area, as well as electrical methods including electrical resistivity tomography (ERT), time domain electro-magnetics (TDEM) and self-potential (SP). These investigations were completed by thermometry, CO2 soil gas measurements, field observations and sampling. This multi-method approach allows geological structures within the caldera to be identified, as well as associated hydrothermal features. The global structure of the caldera is deduced from gravity data, which shows the caldera rim as a high density structure. Large lava fields, emplaced before and after the onset of resurgence, are evidenced by combined gravity, magnetic and resistivity signals. In the middle of the caldera, the Yenkahe dome apparently results from a combination of volcanic and tectonic events, showing that lava extrusion and resurgence have been operating simultaneously or alternately during the Siwi caldera post-collapse history. There is a clear distinction between the western and eastern parts of the dome. The western part is older and records the growth of an initial volcanic cone and the formation of a small caldera. This small caldera (paleo-Yasur caldera), partially filled with lava flows, is the present-day focus of volcanic activity and associated fluid circulation and alteration. The eastern part of the dome is presumably younger, and is characterized by intense, extensive hydrothermal alteration and activity. Its northern part is covered by lava flow piles and exhibits a shallow hydrothermal zone in ERT. The southern part has hydrothermal alteration and activity extending at least down to the base of the resurgent dome. This part of the dome is built up of low cohesion rock and is thus potentially prone to gravitational landslides. Lastly, while self-potential and temperature data suggest that widespread hydrothermal circulation occurs throughout almost all of the caldera, and possibly beyond, the most active parts of this hydrothermal system are associated with the dome. The presence of this active hydrothermal system is the clearest indicator that these methods can provide of a potential shallow magmatic body underneath the dome

Partnerships for innovation: The case of Urban living lab in Turin

Cities are experiencing complex problem such as rapid urbanization, ageing, increased social inequalities, pollution and climate change. Local policymakers are called to handle those challenges with limited resources, increased economic constraints and without the appropriate policy tools. Urban Living Labs can be a useful strategy to deal with multidimensional problems because they engage local actors in experimenting innovative solutions. Urban Living Labs are, in fact, local spaces where municipalities, citizens, and stakeholders define, develop and test innovative products or services, using an open and collaborative approach to innovation, aimed at eliciting knowledge from participants. This chapter investigates how partnerships for ULL are created and managed at the local level, and for the benefit of whom. It analyses the case of the Turin Living Lab created by the Municipality of Turin in 2016 and transformed into Turin City Lab in 2019

Adventive hydrothermal circulation on Stromboli volcano (Aeolian Islands, Italy) revealed by geophysical and geochemical approaches: Implications for general fluid flow models on volcanoes

On March 15th 2007 a paroxysmal explosion occurred at the Stromboli volcano. This event generated a large amount of products,mostly lithic blocks, someofwhich impacted the ground as far as down to 200 m a.s.l., about 1.5 kmfaraway fromthe active vents. Two days after the explosion, a newvapouremissionwas discovered on the north-eastern flank of the volcanic edifice, at 560 m a.s.l., just above the area called “Nel Cannestrà”. This new vapour emission was due to a block impact. In order to investigate the block impact area to understand the appearance of the vapour emission, we conducted on May 2008 a multidisciplinary study involving Electrical Resistivity Tomography (ERT), Ground Penetrating Radar (GPR), Self-Potential (SP), CO2 soil diffuse degassing and soil temperature surveys. This complementary data set revealed the presence of an anomalous conductive body, probably related to a shallow hydrothermal level, at about 10–15 m depth, more or less parallel to the topography. It is the first time that such a hydrothermal fluid flow,with a temperature close to thewater boiling point (76 °C) has been evidenced at Stromboli at this low elevation on the flank of the edifice. The ERT results suggest a possible link between (1) the main central hydrothermal system of Stromboli, located just above the plumbing system feeding the active vents, with a maximum of subsurface soil temperature close to 90 °C and limited by the NeoStromboli summit crater boundary and (2) the investigated area of Nel Cannestrà, at ~500 m a.s.l., a buried eruptive fissure active 9 ka ago. In parallel, SP and CO2 soil diffuse degassingmeasurements suggest in this sector at slightly lower elevation fromthe block impact crater a magmatic and hydrothermal fluid rising system along the N41° regional fault. A complementary ERT profile, on May 2009, carried out from the NeoStromboli crater boundary downto the block impact crater displayed a flank fluid flowapparently connected to a deeper system. The concept of shallow hydrothermal level have been compared to similar ERT results recently obtained onMount Etna and La Fossa cone of Vulcano. This information needs to be taken into account in general fluid flow models on volcanoes. In particular, peripheral thermal waters (as those bordering the northeastern coast of Stromboli) could be contaminated by hydrothermal and magmatic fluids coming from regional faults but also from the summit