Influenza A matrix protein M1 induces lipid membrane deformation via protein multimerization

The matrix protein M1 of the Influenza A virus (IAV) is supposed to mediate viral assembly and budding at the plasma membrane (PM) of infected cells. In order for a new viral particle to form, the PM lipid bilayer has to bend into a vesicle toward the extracellular side. Studies in cellular models have proposed that different viral proteins might be responsible for inducing membrane curvature in this context (including M1), but a clear consensus has not been reached. In the present study, we use a combination of fluorescence microscopy, cryogenic transmission electron microscopy (cryo-TEM), cryo-electron tomography (cryo-ET) and scanning fluorescence correlation spectroscopy (sFCS) to investigate M1-induced membrane deformation in biophysical models of the PM. Our results indicate that M1 is indeed able to cause membrane curvature in lipid bilayers containing negatively charged lipids, in the absence of other viral components. Furthermore, we prove that protein binding is not sufficient to induce membrane restructuring. Rather, it appears that stable M1–M1 interactions and multimer formation are required in order to alter the bilayer three-dimensional structure, through the formation of a protein scaffold. Finally, our results suggest that, in a physiological context, M1-induced membrane deformation might be modulated by the initial bilayer curvature and the lateral organization of membrane components (i.e. the presence of lipid domains)

A Spectroscopic and Cryo-Transmission Electron Microscopy Study

The aggregation behaviour of the cationic pinacyanol chloride in aqueous solution is investigated using absorption and linear dichroism spectroscopies, optical microscopy and cryogenic transmission electron microscopy (cryo-TEM). The investigations are focused on solutions in a concentration range from 50 μM up to 1 mM. At a concentration of 0.7 mM H-aggregates are detected that are characterized by a broad absorption band centred at [similar]511 nm. The aggregates possess a tubular architecture with a single-layer wall thickness of [similar]2.5 nm and an outer diameter of [similar]6.5 nm. Linear dichroism spectroscopy indicates that the molecules are packed with their long axis parallel to the tube axis. These H-aggregates are not stable, but transform into J-aggregates on the time scale of weeks. The kinetics of J-aggregation depends on the dye concentration and the route of sample preparation, but can also be enhanced by shear stress. J-aggregates possess a split absorption spectrum composed of two longitudinally polarized J-bands and one H-band that is polarized perpendicular to the aggregate axis. The J-aggregates are [similar]9 nm wide and several micrometer long fibrils consisting of stacked pairs of ribbons with a dumbbell-shaped density cross-section. Upon aging these ribbons laterally stack face-to-face to form tape-like aggregates

Optical Visualization of Radiative Recombination at Partial Dislocations in GaAs

Individual dislocations in an ultra-pure GaAs epilayer are investigated with spatially and spectrally resolved photoluminescence imaging at 5~K. We find that some dislocations act as strong non-radiative recombination centers, while others are efficient radiative recombination centers. We characterize luminescence bands in GaAs due to dislocations, stacking faults, and pairs of stacking faults. These results indicate that low-temperature, spatially-resolved photoluminescence imaging can be a powerful tool for identifying luminescence bands of extended defects. This mapping could then be used to identify extended defects in other GaAs samples solely based on low-temperature photoluminescence spectra.Comment: 4 pages, 4 figure

Technical Note: A simple method for air–sea gas exchange measurements in mesocosms and its application in carbon budgeting

Mesocosms as large experimental vessels principally provide the opportunity of performing elemental budget calculations e.g. to derive net biological turnover rates. However, the system is in most cases not closed at the water surface and gases can exchange with the atmosphere. Previous attempts to budget carbon pools in mesocosms relied on educated guesses concerning the exchange of CO2 with the atmosphere. Nevertheless, net primary production rates derived from these budget calculations were, despite large uncertainties in air/sea gas exchange, often more reasonable than cumulative extrapolations of bioassays. While bioassays have limitations representing the full spectrum of trophic levels and abiotic conditions inside the mesocosms, calculating dissolved inorganic carbon uptake inside the mesocosms has the potential to deliver net community production rates representative of the enclosed system. Here, we present a simple method for precise determination of air/sea gas exchange velocities in mesocosms using N2O as a deliberate tracer. Beside the application for carbon budgeting, exchange velocities can be used to calculate exchange rates of any gas of known concentration, e.g. to calculate aquatic production rates of climate relevant trace gases. Using an arctic (Kiel Off Shore Mesocosms for future Ocean Simulation) mesocosm experiment as an exemplary dataset, it is shown that application of the presented method largely improves accuracy of carbon budget estimates. Methodology of manipulation, measurement, data processing and conversion to CO2 fluxes are explained. A theoretical discussion of prerequisites for precise gas exchange measurements provides a guideline for the applicability of the method under various experimental conditions

The plasmamembrane calmodulin–dependent calcium pump: a major regulator of nitric oxide synthase I

The plasma membrane calcium/calmodulin-dependent calcium ATPase (PMCA) (Shull, G.E., and J. Greeb. 1988. J. Biol. Chem. 263:8646–8657; Verma, A.K., A.G. Filoteo, D.R. Stanford, E.D. Wieben, J.T. Penniston, E.E. Strehler, R. Fischer, R. Heim, G. Vogel, S. Mathews, et al. 1988. J. Biol. Chem. 263:14152–14159; Carafoli, E. 1997. Basic Res. Cardiol. 92:59–61) has been proposed to be a regulator of calcium homeostasis and signal transduction networks of the cell. However, little is known about its precise mechanisms of action. Knock-out of (mainly neuronal) isoform 2 of the enzyme resulted in hearing loss and balance deficits due to severe inner ear defects, affecting formation and maintenance of otoconia (Kozel, P.J., R.A. Friedman, L.C. Erway, E.N. Yamoah, L.H. Liu, T. Riddle, J.J. Duffy, T. Doetschman, M.L. Miller, E.L. Cardell, and G.E. Shull. 1998. J. Biol. Chem. 273:18693–18696). Here we demonstrate that PMCA 4b is a negative regulator of nitric oxide synthase I (NOS-I, nNOS) in HEK293 embryonic kidney and neuro-2a neuroblastoma cell models. Binding of PMCA 4b to NOS-I was mediated by interaction of the COOH-terminal amino acids of PMCA 4b and the PDZ domain of NOS-I (PDZ: PSD 95/Dlg/ZO-1 protein domain). Increasing expression of wild-type PMCA 4b (but not PMCA mutants unable to bind PDZ domains or devoid of Ca2+-transporting activity) dramatically downregulated NO synthesis from wild-type NOS-I. A NOS-I mutant lacking the PDZ domain was not regulated by PMCA, demonstrating the specific nature of the PMCA–NOS-I interaction. Elucidation of PMCA as an interaction partner and major regulator of NOS-I provides evidence for a new dimension of integration between calcium and NO signaling pathways

Technical Note: The determination of enclosed water volume in large flexible-wall mesocosms "KOSMOS"

The volume of water enclosed inside flexible-wall mesocosm bags is hard to estimate using geometrical calculations and can be strongly variable among bags of the same dimensions. Here we present a method for precise water volume determination in mesocosms using salinity as a tracer. Knowledge of the precise volume of water enclosed allows establishment of exactly planed treatment concentrations and calculation of elemental budgets

A low phase noise cavity transmission self-injection locked laser system for atomic physics experiments

Lasers with high spectral purity are indispensable for optical clocks and coherent manipulation of atomic and molecular qubits for applications such as quantum computing and quantum simulation. Stabilisation of the laser to a reference can provide a narrow linewidth and high spectral purity. However, widely-used diode lasers exhibit fast phase noise that prevents high fidelity qubit manipulation. Here we demonstrate a self-injection locked diode laser system utilizing a medium finesse cavity. The cavity not only provides a stable resonance frequency, but at the same time acts as a low-pass filter for phase noise beyond the cavity linewidth of around 100 kHz, resulting in low phase noise from dc to the injection lock limit. We model the expected laser performance and benchmark it using a single trapped $^{40}$Ca$^{+}$-ion as a spectrum analyser. We show that the fast phase noise of the laser at relevant Fourier frequencies of 100 kHz to >2 MHz is suppressed to a noise floor of between -110 dBc/Hz and -120 dBc/Hz, an improvement of 20 to 30 dB over state-of-the-art Pound-Drever-Hall-stabilized extended-cavity diode lasers. This strong suppression avoids incoherent (spurious) spin flips during manipulation of optical qubits and improves laser-driven gates in using diode lasers with applications in quantum logic spectroscopy, quantum simulation and quantum computation.Comment: 10 pages, 4 figure

Plasma Lens Prototype Progress: Plasma Diagnostics And Particle Tracking For ILC e+ Source

In recent years, the concept of high-gradient, symmetric focusing using active plasma lenses has regained notable attention owing to its potential benefits in terms of compactness and beam dynamics when juxtaposed with traditional focusing elements. An enticing application lies in the optical matching of extensively divergent positrons originating from the undulator-based ILC positron source, thereby enhancing the positron yield in subsequent accelerating structures. Through a collaboration between the University of Hamburg and DESY Hamburg, a scaled-down prototype for this purpose has been conceptualized and fabricated. In this presentation, we provide an overview of the ongoing progress in the development of this prototype. Furthermore, first insights into the development of a particle tracking code especially designed for plasma lenses with implemented Bayes optimization, are given.Comment: TALK PRESENTED AT THE INTERNATIONAL WORKSHOP ON FUTURE LINEAR COLLIDERS (LCWS2023), 15-19 MAY 2023. C23-05-15.3., LCWS23, SLAC, Plasma lens, ILC, 3 page