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At least 289 records · Page 16

Pectin Based Graft Copolymer–ZnO Hybrid Nanocomposite for the Adsorptive Removal of Crystal Violet

In this work we report the comparative study of the dye adsorption behavior of a Pectin-based terpolymer gel, pectin-graft-(poly(2-acrylamido-2-methyl-1-propane sulfonic acid)-co-acrylamide) (Pec-g-poy(AMPS-co-AAm)) and its nanocomposite with ZnO, (Pec-g-poy(AMPS-co-AAm)/ZnO) from aqueous solutions. Both the hydrogel systems have been synthesized using microwave irradiation technique through free radical polymerization. Swelling behavior of the systems was studied in buffer solutions of pH ranging from 1.2 to 13 and maximum swelling was observed in neutral medium. The equilibrium swelling of parent gel was found to be 28.12 g/g whereas it was 16.54 in the case of nanocomposite. Cationic dye, crystal violet, is employed to assess the adsorption efficiency of the gel samples from aqueous solutions. The influence of initial dye concentration was investigated. A maximum adsorption of 329 and 568 mg/g has been observed for the parent gel and the nanocomposite respectively. The presence of ZnO is observed to enhance the adsorption capacity of Pec-g-poy(AMPS-co-AAm) gel considerably. The kinetic studies revealed a second order adsorption process with high rate of initial adsorption in the case of the nanocomposite when compared to the parent gel and the isotherm studies reveals that the adsorption process fits well into Langmuir model. The evaluation of thermodynamic parameters indicated the adsorption process to be exothermic and spontaneous. Desorption studies under acidic condition showed that almost 96% of the adsorbed dye molecules could be desorbed to the stripping solution of pH 1.2 which support the use of these hydrogel systems for the repeated use as adsorbents for cationic dyes.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Recognition of Nucleoside Monophosphate Substrates by Haemophilus influenzae Class C Acid Phosphatase

The e (P4) phosphatase from Haemophilus influenzae functions in a vestigial NAD{sup +} utilization pathway by dephosphorylating nicotinamide mononucleotide to nicotinamide riboside. P4 is also the prototype of class C acid phosphatases (CCAPs), which are nonspecific 5{prime},3{prime}-nucleotidases localized to the bacterial outer membrane. To understand substrate recognition by P4 and other class C phosphatases, we have determined the crystal structures of a substrate-trapping mutant P4 enzyme complexed with nicotinamide mononucleotide, 5{prime}-AMP, 3{prime}-AMP, and 2{prime}-AMP. The structures reveal an anchor-shaped substrate-binding cavity comprising a conserved hydrophobic box that clamps the nucleotide base, a buried phosphoryl binding site, and three solvent-filled pockets that contact the ribose and the hydrogen-bonding edge of the base. The span between the hydrophobic box and the phosphoryl site is optimal for recognizing nucleoside monophosphates, explaining the general preference for this class of substrate. The base makes no hydrogen bonds with the enzyme, consistent with an observed lack of base specificity. Two solvent-filled pockets flanking the ribose are key to the dual recognition of 5{prime}-nucleotides and 3{prime}-nucleotides. These pockets minimize the enzyme's direct interactions with the ribose and provide sufficient space to accommodate 5{prime} substrates in an anti conformation and 3{prime} substrates in a syn conformation. Finally, the structures suggest that class B acid phosphatases and CCAPs share a common strategy for nucleotide recognition.

08 HYDROGEN↗

Mammalian histones facilitate antimicrobial synergy by disrupting the bacterial proton gradient and chromosome organization

First proposed as antimicrobial agents, histones were later recognized for their role in condensing chromosomes. Histone antimicrobial activity has been reported in innate immune responses. However, how histones kill bacteria has remained elusive. The co-localization of histones with antimicrobial peptides (AMPs) in immune cells suggests that histones may be part of a larger antimicrobial mechanism in vivo. Here we report that histone H2A enters E. coli and S. aureus through membrane pores formed by the AMPs LL-37 and magainin-2. H2A enhances AMP-induced pores, depolarizes the bacterial membrane potential, and impairs membrane recovery. Inside the cytoplasm, H2A reorganizes bacterial chromosomal DNA and inhibits global transcription. Whereas bacteria recover from the pore-forming effects of LL-37, the concomitant effects of H2A and LL-37 are irrecoverable. Their combination constitutes a positive feedback loop that exponentially amplifies their antimicrobial activities, causing antimicrobial synergy. More generally, treatment with H2A and the pore-forming antibiotic polymyxin B completely eradicates bacterial growth.

59 BASIC BIOLOGICAL SCIENCES↗

Structures of the DarR transcription regulator reveal unique modes of second messenger and DNA binding

The mycobacterial repressor, DarR, a TetR family regulator (TFR), was the first transcription regulator shown to bind c-di-AMP. However, the molecular basis for this interaction and the mechanism involved in DNA binding by DarR remain unknown. Here we describe DarR-c-di-AMP and DarR-DNA structures and complementary biochemical assays. The DarR-c-di-AMP structure reveals a unique effector binding site for a TFR, located between DarR dimer subunits. Strikingly, we show this motif also binds cAMP. The location of the adenine nucleotide binding site between subunits suggests this interaction may facilitate dimerization and hence DNA binding. Indeed, biochemical assays show cAMP enhances DarR DNA binding. Finally, DarR-DNA structures reveal a distinct TFR DNA-binding mechanism involving two interacting dimers on the DNA. Thus, the combined data unveil a newly described second messenger binding motif and DNA binding mode for this important family of regulators.

59 BASIC BIOLOGICAL SCIENCES↗

Potential Role of Acetyl-CoA Synthetase (acs) and Malate Dehydrogenase (mae) in the Evolution of the Acetate Switch in Bacteria and Archaea

Although many Archaea have AMP-Acs (acetyl-coenzyme A synthetase) and ADP-Acs, the extant methanogenic genus Methanosarcina is the only identified Archaeal genus that can utilize acetate via acetate kinase (Ack) and phosphotransacetylase (Pta). Despite the importance of ack as the potential urkinase in the ASKHA phosphotransferase superfamily, an origin hypothesis does not exist for the acetate kinase in Bacteria, Archaea, or Eukarya. Here we demonstrate that Archaeal AMP-Acs and ADP-Acs contain paralogous ATPase motifs previously identified in Ack, which demonstrate a novel relation between these proteins in Archaea. The identification of ATPase motif conservation and resulting structural features in AMP- and ADP-acetyl-CoA synthetase proteins in this study expand the ASKHA superfamily to include acetyl-CoA synthetase. Additional phylogenetic analysis showed that Pta and MaeB sequences had a common ancestor, and that the Pta lineage within the halophilc archaea was an ancestral lineage. Lastly, these results suggested that divergence of a duplicated maeB within an ancient halophilic, archaeal lineage formed a putative pta ancestor. These results provide a potential scenario for the establishment of the Ack/Pta pathway and provide novel insight into the evolution of acetate metabolism for all three domains of life.

59 BASIC BIOLOGICAL SCIENCES↗

Characterization of the Catalytic and Nucleotide Binding Properties of the α-Kinase Domain of Dictyostelium Myosin-II Heavy Chain Kinase A

The α-kinases are a widely expressed family of serine/threonine protein kinases that exhibit no sequence identity with conventional eukaryotic protein kinases. In this report, we provide new information on the catalytic properties of the α-kinase domain of Dictyostelium myosin-II heavy chain kinase-A (termed A-CAT). Crystallization of A-CAT in the presence of MgATP yielded structures with AMP or adenosine in the catalytic cleft together with a phosphorylated Asp-766 residue. The results show that the β- and α-phosphoryl groups are transferred either directly or indirectly to the catalytically essential Asp-766. Biochemical assays confirmed that A-CAT hydrolyzed ATP, ADP, and AMP with k cat values of 1.9, 0.6, and 0.32 min –1 , respectively, and showed that A-CAT can use ADP to phosphorylate peptides and proteins. Binding assays using fluorescent 2'/3'-O-(N-methylanthraniloyl) analogs of ATP and ADP yielded K d values for ATP, ADP, AMP, and adenosine of 20 ± 3, 60 ± 20, 160 ± 60, and 45 ± 15 μm, respectively. Site-directed mutagenesis showed that Glu-713, Leu-716, and Lys-645, all of which interact with the adenine base, were critical for nucleotide binding. Mutation of the highly conserved Gln-758, which chelates a nucleotide-associated Mg 2+ ion, eliminated catalytic activity, whereas loss of the highly conserved Lys-722 and Arg-592 decreased k cat values for kinase and ATPase activities by 3–6-fold. Mutation of Asp-663 impaired kinase activity to a much greater extent than ATPase, indicating a specific role in peptide substrate binding, whereas mutation of Gln-768 doubled ATPase activity, suggesting that it may act to exclude water from the active site.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

amPEPpy 1.0: a portable and accurate antimicrobial peptide prediction tool

Antimicrobial peptides (AMPs) are promising alternative antimicrobial agents. Currently, however, portable, user-friendly and efficient methods for predicting AMP sequences from genome-scale data are not readily available. In this work, we present amPEPpy, an open-source, multi-threaded command-line application for predicting AMP sequences using a random forest classifier.

59 BASIC BIOLOGICAL SCIENCES↗

Conformational communication mediates the reset step in t6A biosynthesis

Abstract The universally conserved N6-threonylcarbamoyladenosine (t6A) modification of tRNA is essential for translational fidelity. In bacteria, t6A biosynthesis starts with the TsaC/TsaC2-catalyzed synthesis of the intermediate threonylcarbamoyl adenylate (TC–AMP), followed by transfer of the threonylcarbamoyl (TC) moiety to adenine-37 of tRNA by the TC-transfer complex comprised of TsaB, TsaD and TsaE subunits and possessing an ATPase activity required for multi-turnover of the t6A cycle. We report a 2.5-Å crystal structure of the T. maritima TC-transfer complex (TmTsaB2D2E2) bound to Mg2+-ATP in the ATPase site, and substrate analog carboxy-AMP in the TC-transfer site. Site directed mutagenesis results show that residues in the conserved Switch I and Switch II motifs of TsaE mediate the ATP hydrolysis-driven reactivation/reset step of the t6A cycle. Further, SAXS analysis of the TmTsaB2D2-tRNA complex in solution reveals bound tRNA lodged in the TsaE binding cavity, confirming our previous biochemical data. Based on the crystal structure and molecular docking of TC–AMP and adenine-37 in the TC-transfer site, we propose a model for the mechanism of TC transfer by this universal biosynthetic system.

Luthra, Amit↗

Structure of the prolyl-tRNA synthetase from the eukaryotic pathogen Giardia lamblia

The structure of Giardia prolyl-tRNA synthetase cocrystallized with proline and ATP shows evidence for half-of-the-sites activity, leading to a corresponding mixture of reaction substrates and product (prolyl-AMP) in the two active sites of the dimer. The genome of the human intestinal parasite Giardia lamblia contains only a single aminoacyl-tRNA synthetase gene for each amino acid. The Giardia prolyl-tRNA synthetase gene product was originally misidentified as a dual-specificity Pro/Cys enzyme, in part owing to its unexpectedly high off-target activation of cysteine, but is now believed to be a normal representative of the class of archaeal/eukaryotic prolyl-tRNA synthetases. The 2.2 Å resolution crystal structure of the G. lamblia enzyme presented here is thus the first structure determination of a prolyl-tRNA synthetase from a eukaryote. The relative occupancies of substrate (proline) and product (prolyl-AMP) in the active site are consistent with half-of-the-sites reactivity, as is the observed biphasic thermal denaturation curve for the protein in the presence of proline and MgATP. However, no corresponding induced asymmetry is evident in the structure of the protein. No thermal stabilization is observed in the presence of cysteine and ATP. The implied low affinity for the off-target activation product cysteinyl-AMP suggests that translational fidelity in Giardia is aided by the rapid release of misactivated cysteine.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Structural Insights into the Catalytic Mechanism of Escherichia coli Selenophosphate Synthetase

Selenophosphate synthetase (SPS) catalyzes the synthesis of selenophosphate, the selenium donor for the biosynthesis of selenocysteine and 2-selenouridine residues in seleno-tRNA. Selenocysteine, known as the 21st amino acid, is then incorporated into proteins during translation to form selenoproteins which serve a variety of cellular processes. SPS activity is dependent on both Mg{sup 2+} and K{sup +} and uses ATP, selenide, and water to catalyze the formation of AMP, orthophosphate, and selenophosphate. In this reaction, the gamma phosphate of ATP is transferred to the selenide to form selenophosphate, while ADP is hydrolyzed to form orthophosphate and AMP. Most of what is known about the function of SPS has derived from studies investigating Escherichia coli SPS (EcSPS) as a model system. Here we report the crystal structure of the C17S mutant of SPS from E. coli (EcSPS{sup C17S}) in apo form (without ATP bound). EcSPS{sup C17S} crystallizes as a homodimer, which was further characterized by analytical ultracentrifugation experiments. The glycine-rich N-terminal region (residues 1 through 47) was found in the open conformation and was mostly ordered in both structures, with a magnesium cofactor bound at the active site of each monomer involving conserved aspartate residues. Mutating these conserved residues (D51, D68, D91, and D227) along with N87, also found at the active site, to alanine completely abolished AMP production in our activity assays, highlighting their essential role for catalysis in EcSPS. Based on the structural and biochemical analysis of EcSPS reported here and using information obtained from similar studies done with SPS orthologs from Aquifex aeolicus and humans, we propose a catalytic mechanism for EcSPS-mediated selenophosphate synthesis.

59 BASIC BIOLOGICAL SCIENCES↗

Substrate-Specific Reorganization of the Conformational Ensemble of CSK Implicates Novel Modes of Kinase Function

Protein kinases use ATP as a phosphoryl donor for the posttranslational modification of signaling targets. It is generally thought that the binding of this nucleotide induces conformational changes leading to closed, more compact forms of the kinase domain that ideally orient active-site residues for efficient catalysis. The kinase domain is oftentimes flanked by additional ligand binding domains that up- or down-regulate catalytic function. C-terminal Src kinase (Csk) is a multidomain tyrosine kinase that is up-regulated by N-terminal SH2 and SH3 domains. Although the X-ray structure of Csk suggests the enzyme is compact, X-ray scattering studies indicate that the enzyme possesses both compact and open conformational forms in solution. Here, we investigated whether interactions with the ATP analog AMP-PNP and ADP can shift the conformational ensemble of Csk in solution using a combination of small angle x-ray scattering and molecular dynamics simulations. We find that binding of AMP-PNP shifts the ensemble towards more extended rather than more compact conformations. Binding of ADP further shifts the ensemble towards extended conformations, including highly extended conformations not adopted by the apo protein, nor by the AMP-PNP bound protein. These ensembles indicate that any compaction of the kinase domain induced by nucleotide binding does not extend to the overall multi-domain architecture. Instead, assembly of an ATP-bound kinase domain generates further extended forms of Csk that may have relevance for kinase scaffolding and Src regulation in the cell.

Small-angle scattering↗

Managing Aging Effects on Dry Cask Storage Systems for Extended Long-Term Storage and Transportation of Used Fuel

The cancellation of the Yucca Mountain repository program in the United States raises the prospect of extended long-term storage (i.e., >120 years) and deferred transportation of used fuel at operating and decommissioned nuclear power plant sites. Under U.S. federal regulations contained in Title 10 of the Code of Federal Regulations (CFR) 72.42, the initial license term for an Independent Spent Fuel Storage Installation (ISFSI) must not exceed 40 years from the date of issuance. Licenses may be renewed by the U.S. Nuclear Regulatory Commission (NRC) at the expiration of the license term upon application by the licensee for a period not to exceed 40 years. Application for ISFSI license renewals must include the following: (1) Time-limited aging analyses (TLAAs) that demonstrate that structures, systems, and components (SSCs) important to safety will continue to perform their intended function for the requested period of extended operation; and (2) a description of the aging management program (AMP) for management of issues associated with aging that could adversely affect SSCs important to safety. In addition, the application must also include design bases information as documented in the most recent updated final safety analysis report as required by 10 CFR 72.70. Information contained in previous applications, statements, or reports filed with the Commission under the license may be incorporated by reference provided that those references are clear and specific. The NRC has recently issued the Standard Review Plan (SRP) for renewal of used-fuel dry cask storage system (DCSS) licenses and Certificates of Compliance (CoCs), NUREG-1927, under which NRC may renew a specific license or a CoC for a term not to exceed 40 years. Both the license and the CoC renewal applications must contain revised technical requirements and operating conditions (fuel storage, surveillance and maintenance, and other requirements) for the ISFSI and DCSS that address aging effects that could affect the safe storage of the used fuel. The information contained in the license and CoC renewal applications will require NRC review to verify that the aging effects on the SSCs in DCSSs/ ISFSIs are adequately managed for the period of extended operation. To date, all of the ISFSIs located across the United States with more than 1,500 dry casks loaded with used fuel have initial license terms of 20 years; three ISFSIs (Surry, H.B. Robinson and Oconee) have received their renewed licenses for 20 years, and two other ISFSIs (Calvert Cliffs and Prairie Island) have applied for license renewal for 40 years. This report examines issues related to managing aging effects on the SSCs in DCSSs/ISFSIs for extended long-term storage and transportation of used fuels, following an approach similar to that of the Generic Aging Lessons Learned (GALL) report, NUREG-1801, for the aging management and license renewal of nuclear power plants. The report contains five chapters and an appendix on quality assurance for aging management programs for used-fuel dry storage systems. Chapter I of the report provides an overview of the ISFSI license renewal process based on 10 CFR 72 and the guidance provided in NUREG-1927. Chapter II contains definitions and terms for structures and components in DCSSs, materials, environments, aging effects, and aging mechanisms. Chapter III and Chapter IV contain generic TLAAs and AMPs, respectively, that have been developed for managing aging effects on the SSCs important to safety in the dry cask storage system designs described in Chapter V. The summary descriptions and tabulations of evaluations of AMPs and TLAAs for the SSCs that are important to safety in Chapter V include DCSS designs (i.e., NUHOMS{reg_sign}, HI-STORM 100, Transnuclear (TN) metal cask, NAC International S/T storage cask, ventilated storage cask (VSC-24), and the Westinghouse MC-10 metal dry storage cask) that have been and continue to be used by utilities across the country for dry storage of used fuel to date. The goal of this report is to help establish the technical basis for extended long-term storage and transportation of used fuel.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Integrated Predictive Tools for Customizing Microstructure and Material Properties of Additively Manufactured Aerospace Components

Additive Manufacturing (AM) refers to a process by which digital three-dimensional (3-D) design data is converted to build up a component by depositing material layer-by-layer. United Technologies Corporation (UTC) is currently involved in fabrication and certification of several AM aerospace structural components made from aerospace materials. This is accomplished by using optimized process parameters determined through numerous design-of-experiments (DOE)-based studies. Certification of these components is broadly recognized as a significant challenge, with long lead times, very expensive new product development cycles and very high energy consumption. Because of these challenges, United Technologies Research Center (UTRC), together with UTC business units have been developing and validating an advanced physics-based process model. The specific goal is to develop a physics-based framework of an AM process and reliably predict fatigue properties of built-up structures as based on detailed solidification microstructures. Microstructures are predicted using process control parameters including energy source power, scan velocity, deposition pattern, and powder properties. The multi-scale multi-physics model requires solution and coupling of governing physics that will allow prediction of the thermal field and enable solution at the microstructural scale. The state-of-the-art approach to solve these problems requires a huge computational framework and this kind of resource is only available within academia and national laboratories. The project utilized the parallel phase-fields codes at Oak Ridge National Laboratory (ORNL) and Lawrence Livermore National Laboratory (LLNL), along with the high-performance computing (HPC) capabilities existing at the two labs to demonstrate the simulation of multiple dendrite growth in threedimensions (3-D). The LLNL code AMPE was used to implement the UTRC phase field model that was previously developed for a model binary alloy, and the simulation results were compared against the UTRC simulation results, followed by extension of the UTRC model to simulate multiple dendrite growth in 3-D. The ORNL MEUMAPPS code was used to simulate dendritic growth in a model ternary alloy with the same equilibrium solidification range as the Ni-base alloy 718 using realistic model parameters, including thermodynamic integration with a Calphad based model for the ternary alloy. Implementation of the UTRC model in AMPE met with several numerical and parametric issues that were resolved and good comparison between the simulation results obtained by the two codes was demonstrated for two dimensional (2-D) dendrites. 3-D dendrite growth was then demonstrated with the AMPE code using nondimensional parameters obtained in 2-D simulations. Multiple dendrite growth in 2-D and 3-D were demonstrated using ORNL’s MEUMAPPS code using simple thermal boundary conditions. MEUMAPPS was then modified to incorporate the complex, time-dependent thermal boundary conditions obtained by UTRC’s thermal modeling of single track AM experiments to drive the phase field simulations. The results were in good agreement with UTRC’s experimental measurements.

36 MATERIALS SCIENCE↗

Chefornak Energy Configuration Options: Energy Infrastructure Optimization to Reduce Fuel Cost and Dependence in Chefornak, Alaska

The U.S. Department of Energy is funding, through the Grid Modernization Program, the Alaska Microgrid Partnership (AMP), a multi-stakeholder collaborative comprised of national labs and Alaska based partners. The over-arching goal of AMP is to reduce the use of total imported fuel into communities to secure all energy services by at least 50% in Alaska's remote microgrids without increasing system lifecycle costs, while improving overall system reliability, security, and resilience. One goal of AMP is to investigate if a combination of Energy Efficiency (EE) and high contribution (from Renewable Energy (RE)) power systems can reduce total imported energy usage by 50% while lowering Life Cycle Costs (LCC) and improving reliability and resiliency. A techno-economic analysis was conducted for the community of Chefornak to investigate system architectures that could meet the goal of reducing imported fuel consumption by 50% or more.

24 POWER TRANSMISSION AND DISTRIBUTION↗

A Low Cost SIPM Evaluation and Control Prototyping System including accurate bias voltage generation, leakage current measurement, and temperature control using Peltier cooling

We describe an electronics system to operate SIPMs in the laboratory. The system is comprised of a single channel amplifier board (SIPM Amp) and a 4-channel control board to which up to 4 SIPM Amp boards can be connected. The control board supplies a common low voltage power for the amplifiers. All channels feature individually regulated bias and Peltier power voltages, along withPt10K RTD readout. A graphical user interface allows control of the bias voltages, amplifier voltage, Peltier voltages, and also provides for readback of leakage currents and other voltages. The GUI app has a feedback loop to measure the SIPM temperature (with the PT10K) and generate correction voltages to the Peltier to regulate the temperature. The temperatures, which can be achieved by Peltier cooling depend on the thermal design of the setup, the practical limit being about 50C below ambient. The SIPM Amp. board is a small (3x5cm) PCB that has a single SIPM readout channel. The SIPM scan be surface mounted on the pcb or connected to a 2-pin 0.100" socket. The area around the SIPM footprint is thermally isolated by perforations in the PCB. The RTD is located inside the thermal break. Onboard is BV and LV filtering. The 10X amplifier is based on MiniCircuits' GALI-S66+ integrated circuit, tweaked for a 6V power. The amplifier was selected for a combination of high bandwidth, low noise, and low power. An accoupled amplifier output is connected to an SMA connector. For best operation of this inverting amplifier the output signal should be in the range of 0 to -1V.The Amplifier board is connected to the Control board with a 10-conductor 0.025" pitch ribbon cable, carrying all the services. Peltier power for currents more than 1A should use a separate 2-contact connector.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Ageing Management for Extended Long-Term Dry Storage of Spent Nuclear Fuel and Transportation

The principal objectives of the Coordinated Research Project (CRP) and CRADA were to (1) investigate how ageing effects leading to degradation of materials used in the spent fuel dry storage systems could be managed by ageing management programs (AMPs) and (2) using existing AMPs as a basis, establish guidance on how to develop, generate, and maintain AMPs for dry storage systems of spent nuclear fuel (SNF) that can be accomplished in various ways. The PI (Dr. Liu) is an internationally recognized expert on ageing management for license renewal of nuclear power plants and independent spent fuel dry storage installations. Dr. Liu was invited by IAEA to participate in this CRP and served as Chair of the Working Group, supporting the IAEA CRP Lead, with other members from Argentina, the Czech Republic, France, Germany, Hungary, Japan, Pakistan, Spain, Switzerland, United Kingdom, and the United States of America. Argonne’s scope of work in the CRADA for the CRP included (1) ageing management guidance documents developed by Argonne for DOE and used by the Nuclear Regulatory Commission (NRC) and industry and (2) research leveraged from the DOE Office of Nuclear Energy on the mechanical properties of high-burnup fuel cladding and the ARG-US remote monitoring systems technology developed for the DOE Packaging Certification Program, Office of Packaging and Transportation, Office of Environmental Management.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Fermilab 2025 Summer Internship: Repairing Pre-Amplifiers with Mu2e Electronics Installation Team

The author spent nine weeks over summer 2025 working on the tracker electronics installation team for the Mu2e experiment. One of her main responsibilities was repairing high voltage (HV) and calibration (Cal) pre-amplifiers (pre-amps). During installation, the fragile wires connecting the two sockets to the pre-amp board must be bent, often leading to breakage. During production, the sockets and wires were initially soldered to the board at UC Berkely, then the whole pre-amp was coated in parylene before transport to Fermilab. The interns were able to expedite the repairs, and thus whole installation process, by using an alternative method on-site with epoxy. Another task they were responsible for, not included in the original project specifications, was attaching copper clips to specific vias on the Cals to reduce noise. The talk will give listeners insight into the daily problem-solving required by the novel technologies in the Mu2e project. The author would like to acknowledge her fellow Monmouth College undergraduate interns, Lizzie Durfee and Gianna Maughan, advisor and PI of the DOE RENEW Grant Dr. Christopher G. Fasano, and the Mu2e team lead by co-spokesperson Dr. Bob Bernstein and tracker L2 manager Dr. Brendan Kiburg.

de Zwart, Bronte [Monmouth Coll.]↗

Structural Characterization of Maize SIRK1 Kinase Domain Reveals an Unusual Architecture of the Activation Segment

Kinases are primary regulators of plant metabolism and excellent targets for plant breeding. However, most kinases, including the abundant receptor-like kinases (RLK), have no assigned role. SIRK1 is a leucine-rich repeat receptor-like kinase (LRR-RLK), the largest family of RLK. In Arabidopsis thaliana, SIRK1 (AtSIRK1) is phosphorylated after sucrose is resupplied to sucrose-starved seedlings and it modulates the sugar response by phosphorylating several substrates. In maize, the ZmSIRK1 expression is altered in response to drought stress. In neither Arabidopsis nor in maize has the function of SIRK1 been completely elucidated. As a first step toward the biochemical characterization of ZmSIRK1, we obtained its recombinant kinase domain, demonstrated that it binds AMP-PNP, a non-hydrolysable ATP-analog, and solved the structure of ZmSIRK1- AMP-PNP co-crystal. The ZmSIRK1 crystal structure revealed a unique conformation for the activation segment. In an attempt to find inhibitors for ZmSIRK1, we screened a focused small molecule library and identified six compounds that stabilized ZmSIRK1 against thermal melt. ITC analysis confirmed that three of these compounds bound to ZmSIRK1 with low micromolar affinity. Solving the 3D structure of ZmSIRK1-AMP-PNP co-crystal provided information on the molecular mechanism of ZmSIRK1 activity. Furthermore, the identification of small molecules that bind this kinase can serve as initial backbone for development of new potent and selective ZmSIRK1 antagonists.

59 BASIC BIOLOGICAL SCIENCES↗