Structure of a mutated photosystem II complex reveals changes to the hydrogen-bonding network that affect proton egress during O–O bond formation
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Flesher, David A. (ORCID:0000000282704751)↗
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Thiol post-translational modifications modulate allosteric regulation of the OpcA–G6PDH complex through conformational gate control
In cyanobacteria, the redox-sensitive protein OpcA acts as a metabolic switch for G6PDH, enabling rapid adjustment of reducing power generation from glycogen catabolism and thereby precisely regulating carbon flux between anabolic and catabolic pathways. Although redox-sensitive cysteines in OpcA are known to regulate G6PDH, the mechanisms by which redox post-translational modifications (PTMs) on OpcA control G6PDH structure and activity remain unclear. Here, we combine computational modeling with experimental redox proteomics in Synechococcus elongatus PCC 7942 to dissect this mechanism. Experimentally, redox proteome analysis revealed differential redox PTM patterns, particularly on cysteines within the G6PDH-binding site of OpcA. These environmentally sensitive PTM changes at the interface suggest that thiol modifications in this region form a key regulatory node. More broadly, redox proteomics identified site-specific cysteine modifications under light/dark transitions and circadian cycling, linking distinct redox regimes to discrete PTM states. We employed PTM-Psi simulations to show that thiol PTMs near the OpcA–G6PDH interface are critical for allosteric regulation of G6PDH. The thiol PTMs on OpcA affect a putative gate region in G6PDH for substrate ingress and product egress as well as key hydrogen-bond networks within the active site. We infer that PTMs on OpcA tune the conformational landscapes of individual G6PDH subunits toward functionally relevant configurations according to environmental gradients, biasing the enzyme toward catalytically favorable states. Together, our results reveal a molecular mechanism in which thiol PTMs on OpcA modulate G6PDH structure and function through PTM-induced reorganization of conformational dynamics and allosteric communication. These findings demonstrate that PTM-level regulation provides a critical control layer from genotypes to phenotypes that enables cyanobacteria to rapidly adapt to environmental fluctuations through precise metabolic fine-tuning.
Allosteric regulation↗
Probing substrate water access through the O1 channel of Photosystem II by single site mutations and membrane inlet mass spectrometry
Abstract Light-driven water oxidation by photosystem II sustains life on Earth by providing the electrons and protons for the reduction of CO 2 to carbohydrates and the molecular oxygen we breathe. The inorganic core of the oxygen evolving complex is made of the earth-abundant elements manganese, calcium and oxygen (Mn 4 CaO 5 cluster), and is situated in a binding pocket that is connected to the aqueous surrounding via water-filled channels that allow water intake and proton egress. Recent serial crystallography and infrared spectroscopy studies performed with PSII isolated fromThermosynechococcus vestitus(T. vestitus) support that one of these channels, the O1 channel, facilitates water access to the Mn 4 CaO 5 cluster during its S 2 →S 3 and S 3 →S 4 →S 0 state transitions, while a subsequent CryoEM study concluded that this channel is blocked in the cyanobacteriumSynechocystis sp.PCC 6803, questioning the role of the O1 channel in water delivery. Employing site-directed mutagenesis we modified the two O1 channel bottleneck residues D1-E329 and CP43-V410 (T. vestitusnumbering) and probed water access and substrate exchange via time resolved membrane inlet mass spectrometry. Our data demonstrates that water reaches the Mn 4 CaO 5 cluster via the O1 channel in both wildtype and mutant PSII. In addition, the detailed analysis provides functional insight into the intricate protein-water-cofactor network near the Mn 4 CaO 5 cluster that includes the pentameric, near planar ‘water wheel’ of the O1 channel.
Plant Sciences↗
In Vitro Encapsulation of Functionally Active Abiotic Photosensitizers Inside a Bacterial Microcompartment Shell
Bacterial microcompartments (BMCs) are self-assembling, selectively permeable protein shells that encapsulate enzymes to enhance catalytic efficiency of segments of metabolic pathways through means of confinement. The modular nature of BMC shells' structure and assembly enables programming of shell permeability and underscores their promise in biotechnology engineering efforts for applications in industry, medicine, and clean energy. Realizing this potential requires methods for encapsulation of abiotic molecules, which have been developed here for the first time. We report in vitro cargo loading of BMC shells with ruthenium photosensitizers (RuPS) by two approaches-one involving site-specific covalent labeling and the other driven by diffusion, requiring no specific interactions between cargo molecules and shell proteins. The highly stable shells retain encapsulated cargo over 1 week without egress and preserve RuPS photophysical activity. Finally, this study is an important foundation for further work that will converge biological BMC architecture with synthetic chemistry to facilitate biohybrid photocatalysis.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Crossover as Determinant for Safety and Performance Tradeoffs in Proton Exchange Membrane Water Electrolyzers
Hydrogen (H2) crossover is a pressing challenge constraining safe and efficient operation of proton exchange membrane water electrolyzers (PEMWEs) especially amongst strides to employ thinner membranes, which enables improved energy efficiency, and elevated cathode pressures, that reduces the energy burden on downstream compressors. Here, we develop a microstructure-aware multicomponent reactive-transport framework that resolves dissolved and gaseous H2 transport pathways and mechanistically links electrode architecture to crossover related safety and performance. We show that operability is co-governed by the cathode catalyst layer (CCL) and the anode porous transport layer (APTL) which sets the H2 crossover flux and the egress capacity respectively. Elevated Pt/C ratio in the CCL suppresses crossover flux by up to 23% while a higher APTL porosity lowers H2 in O2 fraction by 0.6% in the anode effluent. We condense the findings into (cathode pressure-current density) maps overlaid with safety limits and performance targets and ultimately define two safety-performance unified metrics to gauge the size and quality of the operating window. Given the push towards higher pressure and deeper turndown for renewable integration, this study provides mechanistic design guidance to prevent crossover-induced safety risks while preserving the desired performance.
Electrolysis↗
The Novel Carbapenem, JDB/PQ-1-219, Has Potent Broad Spectrum Activity against Multi-Drug Resistant Acinetobacter baumannii
Carbapenem resistance in Acinetobacter baumannii, driven largely by class D, along with class A and class B β- lactamases, has severely compromised the utility of these last resort antibiotics. As a result, infections caused by such pathogens are characterized by extremely high mortality rates. Here we describe the antimicrobial activity of the novel C5 methyl-substituted carbapenem JDB/PQ-1−219 against multidrug resistant A. baumannii and the mechanism of its interaction with its major carbapenemase, OXA-23. JDB/PQ-1-219 exhibits potent antimicrobial activity against A. baumannii producing various carbapenemases, with MICs that are all in the clinically susceptible range. The compound has unrestricted ingress through porins and avoids egress by efflux pumps, a unique property when compared to all commercial carbapenems. Kinetic experiments demonstrated that unlike for other carbapenems, acylation of OXA-23 by JDB/ PQ-1-219 is monophasic, and mass spectrometry studies showed that this results from the conversion of all enzyme into a reversible tetrahedral intermediate which gradually transitions into the stable acyl-enzyme complex. No deacylation of this complex is observed over a physiologically relevant time period, making JDB/PQ-1−219 an extremely potent inhibitor of OXA-23. Time-resolved crystallography revealed fine details of active site dynamics, leading to complete inhibition of the enzyme. Together, these studies identify JDB/PQ-1-219 as a uniquely effective novel carbapenem with clinically significant levels of activity against multidrug resistant A. baumannii.
Acinetobacter baumannii↗
Experimental Analysis of Advanced Turbine for Supercritical CO2 Power Cycle
An advanced first stage high pressure turbine blade for a supercritical CO2 power cycle is tested in the Big Rig for Aerothermal Stationary Turbine Analysis (BRASTA) annular cascade at the Purdue Experimental Turbine Aerothermal Laboratory (PETAL) high pressure blow down facility alongside a baseline blade for comparison of aerodynamic performance. Both geometries are tested simultaneously using a novel off-axis rig design to allow for the blade geometries to be scaled up to achieve higher Reynolds numbers. The off-axis design necessitates the use of discrete sectors of airfoils, which are designed and additively manufactured in house using an mSLA printer. Printing the blades allows for unique routing of passage for blade surface static pressure taps that contour to the blade shape instead of requiring a straight line view from surface tap to egress, allowing all instrumentation for 15%, 50%, and 85% span to exist in the same passage. A flow conditioning gauze [1] is placed upstream of the blade passages to impart pressure, Mach, and swirl profiles to mimic the rotor relative frame inlet conditions to the blade. Downstream, a sonic valve is used to alter the backpressure to achieve different pressure ratios for different blowdown setpoints. The design of the rig, sonic wheel, and instrumentation are discussed along with the manufacturing and GD&T of the blades with respect to the build-up of this novel experimental apparatus. Initial commissioning data is obtained from inlet total pressure and temperature rakes, blade static pressure taps, and exit total pressure rakes. Oil visualization is performed using a silicone oil mixture containing titanium dioxide and pigmented for contrast of the blade suction side surface with the hub and shroud endwalls. The viscosity of the oil is tuned so that the oil does not fully thin and blow away during the time of the test, and endoscopic cameras are placed in the rig for live monitoring and recording. The preliminary oil visualization and rake profiles are presented, showcasing the operability of this additively manufactured off-axis flow path design.
Tuite, Logan [Purdue University]↗
eCounter: Inline Per-IP Network Monitoring at Millisecond Resolution via eBPF
Scientific data acquisition (SciDAQ) systems are shifting from archive-based workflows to streaming paradigms, where real-time, fine-grained network monitoring becomes essential. While P4-enabled devices offer per-packet in-band observability, they require specialized switches and routers. Host-side tools like Prometheus exporters lack sufficient temporal granularity. To bridge this gap, we present eCounter, a lightweight, hardware-agnostic, inline telemetry agent built on extended Berkeley Packet Filter (eBPF). eCounter captures per-interface ingress and egress traffic, categorized by IP address and protocol, at millisecond to sub-millisecond resolution. In a 100 Gbps environment, it continuously exports up to 3,257 time-series bins per second with only 4% CPU utilization at a 35¿KiB/s data rate. We evaluate eCounter across diverse NIC MTU settings, hook types, CPU architectures and operating systems, and observed negligible impact on concurrent high-throughput streaming applications. Complexity analysis confirms that it can be readily scaled to distributed SciDAQ deployments.
Mei, Xinxin [Computational Sciences and Technology↗
Community Public Mobility Using On-Demand, Low-Speed Electric Vehicles: A Case Study in Downtown St. Louis, Missouri
Legacy fixed route transit systems designed to serve commuters struggle to provide efficient and effective service for short neighborhood trips and for population groups unable to access and egress transit stops using active modes (e.g., elderly, disabled). Neighborhood on-demand transit (ODT) services using low-speed electric vehicles (LSEV) are an innovative technological solution that can help fill this gap in service (e.g., short, high-frequency trips) for diverse populations and trip types. This study evaluated user characteristics and travel behavior for a neighborhood ODT service (using LSEVs) in downtown St. Louis, Missouri using responses from a community survey (n=244), ridership data, and vehicle trajectory information. A comparative analysis between neighborhood ODT, fixed route transit, and transportation network companies (TNC) was also conducted from the perspectives of total travel time, cost, and greenhouse gas emissions. Ultimately, the goal of the analysis was to motivate and inform holistic public mobility systems where different services are optimized to meet specific community needs. Findings indicate that the neighborhood ODT was effective at reaching diverse populations (elderly (20%), lower income (27%), and households with limited access to private vehicles (34%)). ODT reduced total travel time by 32% compared to fixed route transit, produced 2.4 - 4.3 times less greenhouse gas emissions per passenger mile (compared to transit and TNCs), and was more affordable (free to users) than alternative options ($1 for transit, $10-12 for TNCs). Overall satisfaction rates were high, with 80% of respondents rating the service a 4 or 5 out of 5.
29 ENERGY PLANNING, POLICY, AND ECONOMY↗
Transit Survey - Miami-Dade Metrorail - 2009
This Miami-Dade Metrorail Survey obtained ridership characteristics such as origin-destination patterns, trip purpose, and mode of access and egress. The data obtained from this survey was used to update and validate the Southeast Regional Planning Model (SERPM v6.5) and for transportation planning in the region.
32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
Portal Monitoring Considerations for Treaty Verification Applications
Portal monitoring is one method in a suite of options for treaty verification and can assist in verification of treaty accountable items (TAIs). Portal monitor technology spans a wide range of sensors, including radiation detectors, break-beams, or weight sensors. For radiation-detecting portal monitors, they are able to confirm absence / presence of a radiological signature and to track TAI direction of motion. Portal monitoring uses measurements from strategically placed sensors (e.g., radiation detectors) to record the entry or egress of TAIs. The implementation of portal monitoring relies on three key elements: perimeter definition, portal locations, and sensor technology, and can be implemented in either time-bound or continuous operations depending on what is permitted by the Treaty.
98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗
Challenges and Opportunities for Basic Efficiency Measures in Low-Income Homes: A Southeast Alaska Case Study
Juneau, Alaska is the state's capital city and has a renewable energy goal to reach 80% renewable energy for the space heating and transportation sectors by 2045. In practical terms, this indicates a need to electrify both sectors, to take advantage of the inexpensive hydropower available from the local electric utility, Alaska Energy Light & Power. In an effort to complement and enable electrification, researchers examined the feasibility to deploy storm windows via a case study of installing storm windows in two local low-income homes. Newer models of storm windows provide an extra layer of insulation over existing windows, while preserving operability and views. They can also improve comfort and reduce noise. Researchers worked with the regional housing authority and a local builder to install the storm windows and replace inoperable windows in the two houses in 2021, in addition to conducting pre- and post-installation air leakage tests, energy monitoring, and occupant interviews. The team encountered several challenges, including a lack of egress windows, energy data from a wide variety of heating systems, extremely leaky homes, and installation issues such as windows that were not square. These results point to several barriers to widespread deployment of storm windows in the area, but also open the door to opportunities to design deployment programs that improve safety as well as efficiency.
cold climate↗
Evaluating the impacts of Variable Message Signs on Airport Curbside Performance Using Microsimulation
Curbs play a vital role in facilitating vehicle access and egress for individuals at airports. Inefficiently allocating this resource hinders airport accessibility and productivity, resulting in congestion, longer travel times, and increased pollution. As airport demand fluctuates throughout the day and grows over time, airports face intensified curbside pressure. Yet, curb management research is significantly less robust at airports than in urban areas. Given the unbalanced nature of airport demand—riders tend to arrive simultaneously at specific entrances at certain hours—Variable Message Sign (VMS) arises as a cost-effective technology to divert vehicles from congested to underutilized curbs. Still, VMS implementation faces a significant challenge. Historically, airports have managed VMS heuristically and by intuition rather than an evidence-based approach. This research investigates the impacts of implementing VMS on curb performance at airports. By considering different driver compliance rates (DCR), we aim to determine when the sign should be turned on and off to diverge traffic to avoid undesired externalities while enhancing curb performance. Using a validated agent-based microsimulation model, VISSIM, we analyzed the Seattle-Tacoma (SeaTac) Airport as a case study. We modeled sixteen VMS management scenarios and a baseline where the message sign is not displayed, diverging vehicles between the departures and arrivals access levels at four different moments (early morning, morning, afternoon, and late night). We quantified the effects of VMS using seven metrics, including curb productivity index (CPI), curb accessibility (CA), queue length, queue duration, delay, vehicle counts, and emissions. The results of each scenario were compared against the baseline using absolute and relative changes and Repeated Measures ANOVA. Overall, VMS improved curb performance and traffic conditions at the airport, reducing emissions by 14.8% to 8.9%. Moreover, significant reductions in queue length (1,150 ft to 100 ft) and duration (15 to 144 minutes) were observed in the sending link under all VMS policies. However, impacts on the receiving link varied based on congestion, with significant increases in queue duration (9.8 to 24 min) when congested but no substantial changes in free flow. Notably, diverging vehicles to congested links resulted in non-significant results, and activating late and deactivating late VMS affected curb productivity (-5.8% to -61.4%), curb accessibility (-16.5% to -25.8%), cumulative counts (-33.4% to -59.4%), and vehicle delay (95.98% to 594.3%). Activating VMS before congestion begins in the sending link and deactivating before a queue forms in the receiving link yield the most significant improvements: 8.1% to 10.1% in CPI, 9.4% to9.6% in CA, -29.3% to -77.9% in total delay, -11.6% to -13.9% in total emissions, and 101% to 103% in cumulative counts. As the analysis was made with a wide range of time periods, access levels, driver compliance rates, and scenarios, we believe our findings can provide valuable insights into how airports should manage VMS. Our work introduces a novel approach to the scientific airport literature, as some of our metrics were previously unexplored. Additionally, we propose a methodology that other airports can adopt to maximize their curb performance.
Gutierrez, Jorge D.↗