Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography.

We introduce the spitrobot, a protein crystal plunger, enabling reaction quenching via cryo-trapping with a time-resolution in the millisecond range. Protein crystals are mounted on canonical micromeshes on an electropneumatic piston, where the crystals are kept in a humidity and temperature-controlled environment, then reactions are initiated via the liquid application method (LAMA) and plunging into liquid nitrogen is initiated after an electronically set delay time to cryo-trap intermediate states. High-magnification images are automatically recorded before and after droplet deposition, prior to plunging. The SPINE-standard sample holder is directly plunged into a storage puck, enabling compatibility with high-throughput infrastructure. Here we demonstrate binding of glucose and 2,3-butanediol in microcrystals of xylose isomerase, and of avibactam and ampicillin in microcrystals of the extended spectrum beta-lactamase CTX-M-14. We also trap reaction intermediates and conformational changes in macroscopic crystals of tryptophan synthase to demonstrate that the spitrobot enables insight into catalytic events.

Molecular basis for the allosteric activation mechanism of the heterodimeric imidazole glycerol phosphate synthase complex.

Imidazole glycerol phosphate synthase (HisFH) is a heterodimeric bienzyme complex operating at a central branch point of metabolism. HisFH is responsible for the HisH-catalyzed hydrolysis of glutamine to glutamate and ammonia, which is then used for a cyclase reaction by HisF. The HisFH complex is allosterically regulated but the underlying mechanism is not well understood. Here, we elucidate the molecular basis of the long range, allosteric activation of HisFH. We establish that the catalytically active HisFH conformation is only formed when the substrates of both HisH and HisF are bound. We show that in this conformation an oxyanion hole in the HisH active site is established, which rationalizes the observed 4500-fold allosteric activation compared to the inactive conformation. In solution, the inactive and active conformations are in a dynamic equilibrium and the HisFH turnover rates correlate with the population of the active conformation, which is in accordance with the ensemble model of allostery.

Binary Electronic Synapses for Integrating Digital and Neuromorphic Computation in a Single Physical Platform.

As a promising advanced computation technology, the integration of digital computation with neuromorphic computation into a single physical platform holds the advantage of a precise, deterministic, fast data process as well as the advantage of a flexible, paralleled, fault-tolerant data process. Even though two-terminal memristive devices have been respectively proved as leading electronic elements for digital computation and neuromorphic computation, it is difficult to steadily maintain both sudden-state-change and gradual-state-change in a single device due to the entirely different operating mechanisms. In this work, we developed a digital-analog compatible memristive device, namely, binary electronic synapse, through realizing controllable cation drift in a memristive layer. The devices feature nonvolatile binary memory as well as artificial neuromorphic plasticity with high operation endurance. With strong nonlinearity in switching dynamics, binary switching, neuromorphic plasticity, two-dimension information store, and trainable memory can be implemented by a single device.

Biosynaptic devices based on chicken egg albumen:graphene quantum dot nanocomposites.

Biosynaptic devices based on chicken egg albumen (CEA):graphene quantum dot (GQD) hybrid nanocomposites were fabricated to achieve stable synaptic behaviors. Current-voltage (I-V) curves for the biosynaptic devices under consecutive negative and positive voltage sweeps showed clockwise pinched hysteresis, which is a critical feature of a biological synapse. The effect of the GQD concentration in the CEA layer on the device performance was studied. The retention time of the biosynaptic devices was relatively constant, maintaining a value above 104 s under ambient conditions. The carrier transport mechanisms of the biosynaptic devices were described and analyzed on the basis of the slopes of the I-V curves and their fittings.

Self-Powered Tactile Sensor with Learning and Memory.

Fabrication of human-like intelligent tactile sensors is an intriguing challenge for developing human-machine interfaces. As inspired by somatosensory signal generation and neuroplasticity-based signal processing, intelligent neuromorphic tactile sensors with learning and memory based on the principle of a triboelectric nanogenerator are demonstrated. The tactile sensors can actively produce signals with various amplitudes on the basis of the history of pressure stimulations because of their capacity to mimic neuromorphic functions of synaptic potentiation and memory. The time over which these tactile sensors can retain the memorized information is alterable, enabling cascaded devices to have a multilevel forgetting process and to memorize a rich amount of information. Furthermore, smart fingers by using the tactile sensors are constructed to record a rich amount of information related to the fingers' current actions and previous actions. This intelligent active tactile sensor can be used as a functional element for artificial intelligence.

Highly flexible and stable resistive switching devices based on WS2 nanosheets:poly(methylmethacrylate) nanocomposites.

This paper reports data for the electrical characteristics and the operating mechanisms of flexible resistive switching devices based on WS2 nanosheets (NSs) dispersed in a poly(methyl methacrylate) (PMMA) layer. The ON/OFF ratio of the memristive device based on an Al/WS2 NSs:PMMA/indium tin oxides (ITO) structure was approximately 5.9 × 104. The memristive device based on the WS2 NSs also exhibited the bipolar switching characteristics with low power consumption and great performance in the bent state with radii of the curvatures of 20 and 10 mm. Especially, the results obtained after bending the device were similar to those observed before bending. The device showed nearly the same ON/OFF ratio for a retention time of 1 × 104 sec, and the number of endurance cycles was greater than 1 × 102. The set voltage and the reset voltage probability distributions for the setting and the resetting processes indicated bipolar switching characteristics. The operating and the carrier transport mechanisms of the Al/WS2 NSs:PMMA/ITO device could be explained based on the current-voltage results with the aid of an energy band diagram.

Flexible organic synaptic device based on poly (methyl methacrylate):CdSe/CdZnS quantum-dot nanocomposites.

A synaptic device that functionally mimics a biological synapse is a promising candidate for use as an electronic element in a neuromorphic system. In this study, flexible electronic synaptic devices based on poly (methyl methacrylate) (PMMA):CdSe/CdZnS core-shell quantum-dot (QD) nanocomposites are demonstrated. The current-voltage characteristics for the synaptic devices under consecutive voltage sweeps show clockwise hysteresis, which is a critical feature of an artificial synaptic device. The effect of the CdSe/CdZnS QD concentration on the device performance is studied. The flexible electronic synaptic devices under bending show the similar and stable electrical performances. The memory retention measurements show that the e-synapse exhibits long-term potentiation and depression. The carrier transport mechanisms are analyzed, and thermionic emission and space-charge-limited-current conduction are found to be dominant.

Flexible memristive devices based on polyimide:mica nanosheet nanocomposites with an embedded PEDOT:PSS layer.

Flexible memristive devices with a structure of Al/polyimide:mica/poly(3,4-ethylenedioxythiophene) polystyrene sulfonate/indium-tin-oxide/polyethylene glycol naphthalate showed electrical bistability characteristics. The maximum current margin of the devices with mica nanosheets was much larger than that of the devices without mica nanosheets. For these devices, the current vs. time curves showed nonvolatile characteristics with a retention time of more than 1 × 104 s, and the current vs. number-of-cycles curves demonstrated an endurance for high resistance state/low resistance state switchings of 1 × 102 cycles. As to the operation performance, the "reset" voltage was distributed between 2.5 and 3 V, and the "set" voltage was distributed between -0.7 and -0.5 V, indicative of high uniformity. The electrical characteristics of the devices after full bendings with various radii of curvature were similar to those before bending, which was indicative of devices having ultra-flexibility. The carrier transport and the operation mechanisms of the devices were explained based on the current vs. voltage curves and the energy band diagrams.

Highly-stable write-once-read-many-times switching behaviors of 1D-1R memristive devices based on graphene quantum dot nanocomposites.

One diode and one resistor (1D-1R) memristive devices based on inorganic Schottky diodes and poly(methylsilsesquioxane) (PMSSQ):graphene quantum dot (GQD) hybrid nanocomposites were fabricated to achieve stable memory characteristics. Current-voltage (I-V) curves for the Al/PMSSQ:GQDs/Al/p-Si/Al devices at room temperature exhibited write-once, read-many-times memory (WORM) characteristics with an ON/OFF ratio of as large as 104 resulting from the formation of a 1D-1R structure. I-V characteristics of the WORM 1D-1R device demonstrated that the memory and the diode behaviors of the 1D-1R device functioned simultaneously. The retention time of the WORM 1D-1R devices could be maintained at a value larger than 104 s under ambient conditions. The operating mechanisms of the devices were analyzed on the basis of the I-V results and with the aid of the energy band diagram.

ATP-dependent DNA binding, unwinding, and resection by the Mre11/Rad50 complex.

ATP-dependent DNA end recognition and nucleolytic processing are central functions of the Mre11/Rad50 (MR) complex in DNA double-strand break repair. However, it is still unclear how ATP binding and hydrolysis primes the MR function and regulates repair pathway choice in cells. Here,Methanococcus jannaschii MR-ATPγS-DNA structure reveals that the partly deformed DNA runs symmetrically across central groove between two ATPγS-bound Rad50 nucleotide-binding domains. Duplex DNA cannot access the Mre11 active site in the ATP-free full-length MR complex. ATP hydrolysis drives rotation of the nucleotide-binding domain and induces the DNA melting so that the substrate DNA can access Mre11. Our findings suggest that the ATP hydrolysis-driven conformational changes in both DNA and the MR complex coordinate the melting and endonuclease activity.

DNA end recognition by the Mre11 nuclease dimer: insights into resection and repair of damaged DNA.

The Mre11-Rad50-Nbs1 (MRN) complex plays important roles in sensing DNA damage, as well as in resecting and tethering DNA ends, and thus participates in double-strand break repair. An earlier structure of Mre11 bound to a short duplex DNA molecule suggested that each Mre11 in a dimer recognizes one DNA duplex to bridge two DNA ends at a short distance. Here, we provide an alternative DNA recognition model based on the structures of Methanococcus jannaschii Mre11 (MjMre11) bound to longer DNA molecules, which may more accurately reflect a broken chromosome. An extended stretch of B-form DNA asymmetrically runs across the whole dimer, with each end of this DNA molecule being recognized by an individual Mre11 monomer. DNA binding induces rigid-body rotation of the Mre11 dimer, which could facilitate melting of the DNA end and its juxtaposition to an active site of Mre11. The identified Mre11 interface binding DNA duplex ends is structurally conserved and shown to functionally contribute to efficient resection, non-homologous end joining, and tolerance to DNA-damaging agents when other resection enzymes are absent. Together, the structural, biochemical, and genetic findings presented here offer new insights into how Mre11 recognizes damaged DNA and facilitates DNA repair.

Highly conductive and transparent Ag honeycomb mesh fabricated using a monolayer of polystyrene spheres.

We describe the design principles and fabrication of Ag honeycomb mesh as a transparent conductive electrode using a polystyrene (PS) sphere template. Monolayers of PS spheres with different diameters, such as 600 nm, 3 µm, and 10 µm, are studied as templates to form Ag mesh with high transmittance. Since the parasitic Ag islands degrade the transmittance, both heat pretreatment and wet etching are used to control the area covered by parasitic Ag islands. The trade-off between transmittance and conductivity forces us to use larger diameter PS spheres. Ten-micron PS spheres are chosen as the template for the PS sphere monolayer, and heat pretreatment and Ag wet etching are used to demonstrate that the Ag honeycomb mesh transparent electrodes have high performance. The transmittance and the sheet resistance are 83% and 20 Ω/sq, which are comparable to commercial ITO electrodes.