SSM-based electrophysiology, a label-free real-time method reveals sugar binding & transport events in SGLT1.

Here, we present a solid-supported membrane (SSM)-based electrophysiological approach to study sugar binding and Na+/glucose cotransport by SGLT1 in membrane vesicles. SSM-based electrophysiology delivers a cumulative real-time current readout from numerous SGLT1 proteins simultaneously using a gold-coated sensor chip. In contrast to conventional techniques, which mainly operate with voltage steps, currents are triggered by sugar or sodium addition. Sugar concentration jumps in the presence of sodium lead to transport currents between 5 and 10 nA. Remarkably, in the absence of sodium (i.e. no transport), we observed fast pre-steady-state (PSS) currents with time constants between 3 and 10 ms. These PSS currents mainly originate from sugar binding. Sodium binding does not induce PSS currents. Due to high time resolution, PSS currents were distinguished from transport and eventually correlated with conformational transitions within the sugar translocation pathway. In addition, we analyzed the impact of driving forces on transport and binding currents, showing that membrane voltage and sodium concentration gradients lead to an increased transport rate without affecting sugar binding kinetics. We also compared Na+/sugar efflux with physiologically relevant influx and found similar transport rates, but lower affinity in efflux mode. SSM-based electrophysiology is a powerful technique, which overcomes bottlenecks for transport measurements observed in other techniques such as the requirement of labels or the lack of real-time data. Rapid solution exchange enables the observation of substrate-induced electrogenic events like conformational transitions, opening novel perspectives for in-depth functional studies of SGLT1 and other transporters.

Low-Power Laser Ignition of an Antenna-Type Secondary Energetic Copper Complex: Synthesis, Characterization, Evaluation, and Ignition Mechanism Studies.

In recent years, development of new energetic compounds and formulations, suitable for ignition with relatively low-power lasers, is a highly active and competitive field of research. The main goal of these efforts is focused on achieving and providing much safer solutions for various detonator and initiator systems. In this work, we prepared, characterized, and studied thermal and ignition properties of a new laser-ignitable compound, based on the 5,6-bis(ethylnitroamino)-N'2,N'3-dihydroxypyrazine-2,3-bis(carboximidamide) (DS3) proligand. This new energetic proligand was prepared in three steps, starting with 5,6-bis(ethylamino)-pyrazine-2,3-dicarbonitrile. Crystallography studies of the DS3-derived Cu(II) complex (DS4) revealed a unique stacked antenna-type structure of the latter compound. DS4 has an exothermal temperature of 154.5 °C and was calculated to exhibit a velocity of detonation of 6.36 km·s-1 and a detonation pressure of 15.21 GPa. DS4 showed properties of a secondary explosive, having sensitivity to impact, friction, and electrostatic discharge of 8 J, 360 N, and 12 mJ, respectively. In order to study the mechanism of ignition by a laser (using a diode laser, 915 nm), we conducted a set of experiments that enabled us to characterize a photothermal ignition mechanism. Furthermore, we found that a single pulse, with a time duration of 1 ms and with a total energy of 4.6 mJ, was sufficient for achieving a consistent and full ignition of DS4. Dual-pulse experiments, with variable time intervals between the laser pulses, showed that DS4 undergoes ignition via a photothermal mechanism. Finally, calculating the chemical mechanism of the formation of the complex DS4 and modeling its anhydrous and hydrated crystal structures (density functional theory calculations using Gaussian and HASEM software) allowed us to pinpoint a more precise location of water molecules in experimental crystallographic data. These results suggest that DS4 has potential for further development to a higher technology readiness level and for integration into small-size safe detonator systems as for many civil, aerospace, and defense applications.