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At least 271 records · Page 15

Structural analysis of a new carotenoid-binding protein: the C-terminal domain homolog of the OCP

Abstract The Orange Carotenoid Protein (OCP) is a water-soluble protein that governs photoprotection in many cyanobacteria. The 35 kDa OCP is structurally and functionally modular, consisting of an N-terminal effector domain (NTD) and a C-terminal regulatory domain (CTD); a carotenoid spans the two domains. The CTD is a member of the ubiquitous Nuclear Transport Factor-2 (NTF2) superfamily (pfam02136). With the increasing availability of cyanobacterial genomes, bioinformatic analysis has revealed the existence of a new family of proteins, homologs to the CTD, the C-terminal domain-like carotenoid proteins (CCPs). Here we purify holo-CCP2 directly from cyanobacteria and establish that it natively binds canthaxanthin (CAN). We use small-angle X-ray scattering (SAXS) to characterize the structure of this carotenoprotein in two distinct oligomeric states. A single carotenoid molecule spans the two CCPs in the dimer. Our analysis with X-ray footprinting-mass spectrometry (XFMS) identifies critical residues for carotenoid binding that likely contribute to the extreme red shift (ca. 80 nm) of the absorption maximum of the carotenoid bound by the CCP2 dimer and a further 10 nm shift in the tetramer form. These data provide the first structural description of carotenoid binding by a protein consisting of only an NTF2 domain.

59 BASIC BIOLOGICAL SCIENCES↗

Modular droplet injector for sample conservation providing new structural insight for the conformational heterogeneity in the disease-associated NQO1 enzyme

Droplet injection strategies are a promising tool to reduce the large amount of sample consumed in serial femtosecond crystallography (SFX) measurements at X-ray free electron lasers (XFELs) with continuous injection approaches. Here, we demonstrate a new modular microfluidic droplet injector (MDI) design that was successfully applied to deliver microcrystals of the human NAD(P)H:quinone oxidoreductase 1 (NQO1) and phycocyanin. We investigated droplet generation conditions through electrical stimulation for both protein samples and implemented hardware and software components for optimized crystal injection at the Macromolecular Femtosecond Crystallography (MFX) instrument at the Stanford Linac Coherent Light Source (LCLS). Under optimized droplet injection conditions, we demonstrate that up to 4-fold sample consumption savings can be achieved with the droplet injector. In addition, we collected a full data set with droplet injection for NQO1 protein crystals with a resolution up to 2.7 Å, leading to the first room-temperature structure of NQO1 at an XFEL. NQO1 is a flavoenzyme associated with cancer, Alzheimer's and Parkinson's disease, making it an attractive target for drug discovery. Further, our results reveal for the first time that residues Tyr128 and Phe232, which play key roles in the function of the protein, show an unexpected conformational heterogeneity at room temperature within the crystals. These results suggest that different substates exist in the conformational ensemble of NQO1 with functional and mechanistic implications for the enzyme's negative cooperativity through a conformational selection mechanism. Our study thus demonstrates that microfluidic droplet injection constitutes a robust sample-conserving injection method for SFX studies on protein crystals that are difficult to obtain in amounts necessary for continuous injection, including the large sample quantities required for time-resolved mix-and-inject studies.

59 BASIC BIOLOGICAL SCIENCES↗

Structure, function and evolution of the cyanobacterial orange carotenoid protein and its homologs

The orange carotenoid protein (OCP) is a water-soluble, photoactive protein involved in thermal dissipation of excess energy absorbed by the light-harvesting phycobilisomes (PBS) in cyanobacteria. The OCP is structurally and functionally modular, consisting of a sensor domain, an effector domain and a keto-carotenoid. On photoactivation, the OCP converts from a stable orange form, OCP O , to a red form, OCP R . Activation is accompanied by a translocation of the carotenoid deeper into the effector domain. The increasing availability of cyanobacterial genomes has enabled the identification of new OCP families (OCP1, OCP2, OCPX). The fluorescence recovery protein (FRP) detaches OCP1 from the PBS core, accelerating its back-conversion to OCP O ; by contrast, other OCP families are not regulated by FRP. N-terminal domain homologs, the helical carotenoid proteins (HCPs), have been found among diverse cyanobacteria, occurring as multiple paralogous groups, with two representatives exhibiting strong singlet oxygen ( 1 O 2 ) quenching (HCP2, HCP3) and another capable of dissipating PBS excitation (HCP4). Crystal structures are presently available for OCP1 and HCP1, and models of other HCP subtypes can be readily produced as a result of strong sequence conservation, providing new insights into the determinants of carotenoid binding and 1 O 2 quenching

59 BASIC BIOLOGICAL SCIENCES↗

Idaho National Laboratory’s Mobile Hot Cell Transportation: Engineering Solutions for Global Disused Sealed Radioactive Sources.

Title: Idaho National Laboratory’s Mobile Hot Cell Transportation: Engineering Solutions for Global Disused Sealed Radioactive Sources. Abstract: The Mobile Hot Cell (MHC), currently under development by Idaho National Laboratory (INL) for the Off-Site Source Recovery Project (OSRP), is designed to help international partners meet the unique challenges of end-of-life radioactive material management. The MHC will provide a critical resource for countries that require assistance securing and disposing of Disused Sealed Radioactive Sources (DSRS) and orphaned sources in challenging environments, allowing these sources to be secured against misuse and nefarious activities. The MHC is a rapidly deployable system for conditioning and preparing end-of-life radioactive sources for transportation or storage. It is designed to handle sources of up to 1,000 Ci Co-60 equivalent while maintaining full radiological and biological containment. It will be deployable within 48 hours of an alert, making it ideal for emergency situations. The MHC features an operational suite for control, support racks for electronics, pneumatics, and welding systems, and a modular robust steel structure providing radiological shielding and internal robotic support. This design allows configurations for multiple device types to be conditioned and the ability to safely manage routine issues such as leaking or damaged sources. The MHC has been designed with the transportation challenges of rapid deployment to difficult environments in mind. The system weighs approximately 150,000 pounds, with individual systems breaking down into pieces not exceeding 20,000 pounds. Components are to be transportable on standard 20ft ISO containers, with shielding shells on 20ft flat racks. It is estimated that a total of eight containers and flat racks will be required. The use of 20ft containers, as opposed to 40ft containers, minimizes the impact on less developed road infrastructures, enabling the MHC to be positioned in constrained environments such as hospital parking lots. The system’s modularity also allows for deployment using smaller equipment, such as a 10-ton boom truck or forklift, which is crucial given the potential logistical challenges in different countries. This transportation strategy, evaluated in collaboration with Utah State University, ensures the MHC can be deployed via ground, rail, sea, or air, addressing the primary concern of international transport logistics.

99 - GENERAL AND MISCELLANEOUS↗

A Computer Program to Generate Tri-Truss Structures for On- and Off-Axis Telescope Dishes

The size of traditional space telescopes has been limited by the size of the launch vehicle shroud size. Designs of space telescopes with larger apertures and greater resolving power can be achieved with multiple launches incorporating modular elements and in-space assembly techniques. The modular elements of an in-space assembled telescope include segmented reflectors supported on an assembled truss structure. The modular truss structures are most commonly based on a tessellation of triangles across a reflector’s surface, and in some cases utilize deployable trusses to simplify in-space assembly. One design effort, the in-Space Assembled Telescope (iSAT), proposes the use of deployable “tri-truss” modules to form an off-set parabolic reflector. In support of the iSAT, a computer program was developed to tesselate tri-truss modules over a parabolic surface. Based on several user-defined variables, the program renders a tri-truss structure and generates a stereolithography file of the generated truss structure for three dimensional (3D) printing. In addition, due to the importance of packing efficiency for space launch, the program attempts to minimize unnecessary differences in strut lengths using a grade parameter based on previous work.

Structures↗

TriTruss Packaging and Deployment Trade Study

An architecture and feasibility study of next generation In-Space Assembled Telescopes (iSAT) concluded that robotic in-space assembly of modular components is necessary to enable large (>15-meter diameter) primary apertures. The iSAT study recommended that the foundational structure be assembled from modular TriTruss modules that are packaged for launch, deployed on-orbit and robotically assembled into the final configuration. This paper will describe and summarize the results of a trade study that investigated viable concept of operations (ConOps) for the packaging and deployment (P&D) of individual planar TriTruss modules that could be assembled to form a large aperture iSAT. The ideal TriTruss P&D concept is defined as one that: allows for efficient packaging; has sufficient geometric versatility to be launch vehicle independent; provides a stiff and lightweight structure; has low mechanical complexity; and has component modularity. The P&D concept should allow for prelaunch subsystem or utility integration if required. The concept would be kinematically simple and be robotically deployed using a minimum number of specialized tools. In this first phase of an ongoing more comprehensive trade study, concepts were proposed and then evaluated based on initial metrics representing features of an ideal P&D concept. The P&D concepts evaluated are categorized as: core collapse, face collapse, and erectable structures. Sub-scale models were constructed to help understand the kinematics and mechanical complexity required to enable P&D. Based on a weighting scale, the most promising candidate P&D concepts have selected and will undergo more rigorous structural design, analysis, and testing in the study’s next phase. The ultimate goal of the comprehensive trade study will be to recommend a single TriTruss design and associated P&D concept that will be built and evaluated at NASA Langley Research Center’s In-Space Assembly Laboratory.

in-space assembly (ISA)↗

Triadic Closure in Configuration Models with Unbounded Degree Fluctuations

The configuration model generates random graphs with any given degree distribution, and thus serves as a null model for scale-free networks with power-law degrees and unbounded degree fluctuations. For this setting, we study the local clustering c(k), i.e., the probability that two neighbors of a degree-k node are neighbors themselves. We show that c(k) progressively falls off with k and the graph size n and eventually for k=Ω(√n) settles on a power law c(k)∼n{sup 5−2τ}k{sup −2(3−τ)} with τ∈(2,3) the power-law exponent of the degree distribution. This fall-off has been observed in the majority of real-world networks and signals the presence of modular or hierarchical structure. Our results agree with recent results for the hidden-variable model and also give the expected number of triangles in the configuration model when counting triangles only once despite the presence of multi-edges. We show that only triangles consisting of triplets with uniquely specified degrees contribute to the triangle counting.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Synthetic OCP heterodimers are photoactive and recapitulate the fusion of two primitive carotenoproteins in the evolution of cyanobacterial photoprotection

The orange carotenoid protein (OCP) governs photoprotection in the majority of cyanobacteria. It is structurally and functionally modular, comprised of a C-terminal regulatory domain (CTD), an N-terminal effector domain (NTD) and a ketocarotenoid; the chromophore spans the two domains in the ground state and translocates fully into the NTD upon illumination. Using both the canonical OCP1 from Fremyella diplosiphon and the presumably more primitive OCP2 paralog from the same organism, we show that an NTD-CTD heterodimer forms when the domains are expressed as separate polypeptides. The carotenoid is required for the heterodimeric association, assembling an orange complex which is stable in the dark. Both OCP1 and OCP2 heterodimers are photoactive, undergoing light-driven heterodimer dissociation, but differ in their ability to reassociate in darkness, setting the stage for bioengineering photoprotection in cyanobacteria as well as for developing new photoswitches for biotechnology. Additionally, we reveal that homodimeric CTD can bind carotenoid in the absence of NTD, and name this truncated variant the C-terminal domain-like carotenoid protein (CCP). This finding supports the hypothesis that the OCP evolved from an ancient fusion event between genes for two different carotenoid-binding proteins ancestral to the NTD and CTD. We suggest that the CCP and its homologs constitute a new family of carotenoproteins within the NTF2-like superfamily found across all kingdoms of life.

59 BASIC BIOLOGICAL SCIENCES↗

Pressio

Reduced-order Models (ROMs). ROMs are "compressed" approximations of high-fidelity computational models. Implementing ROM methods in production codes is typically intrusive, i.e. the native codes must be adapted and modified, with individual ROM methods reimplemented in every application code. This clearly constitutes an obstacle to users who want to use ROMs. To address this, we have developed Pressio, which is a C++ header-only ROM library that can be used by an application code in a non-intrusive fashion. The main objectives behind the design of Pressio are: (a) support for general computational models (data structures, PDE types); (b) heterogeneous hardware (CPU, GPU); (c) performance; and (d) extensibility. Using metaprogramming techniques, Pressio detects and leverages the application?s native linear algebra data structures (e.g. vector and matrix) to run the ROM methods. The application only needs to expose the velocity vector and Jacobian matrix via an adapter class. Pressio checks at compile time that the target application meets the correct API. The library is structured in several modular packages including (a) non-linear least-square solvers (e.g. Gauss-Newton with and without line-search); (b) singular value decomposition (for the computation of the ROM generalized coordinates) and (c) algebraic hyper-reduction.SAND2019-5878 M

Blonigan, Patrick↗

Pressio v.1.0

Reduced-order Models (ROMs). ROMs are "compressed" approximations of high-fidelity computational models. Implementing ROM methods in production codes is typically intrusive, i.e. the native codes must be adapted and modified, with individual ROM methods reimplemented in every application code. This clearly constitutes an obstacle to users who want to use ROMs. To address this, we have developed Pressio, which is a C++ header-only ROM library that can be used by an application code in a non-intrusive fashion. The main objectives behind the design of Pressio are: (a) support for general computational models (data structures, PDE types); (b) heterogeneous hardware (CPU, GPU); (c) performance; and (d) extensibility. Using metaprogramming techniques, Pressio detects and leverages the application?s native linear algebra data structures (e.g. vector and matrix) to run the ROM methods. The application only needs to expose the velocity vector and Jacobian matrix via an adapter class. Pressio checks at compile time that the target application meets the correct API. The library is structured in several modular packages including (a) non-linear least-square solvers (e.g. Gauss-Newton with and without line-search); (b) singular value decomposition (for the computation of the ROM generalized coordinates) and (c) algebraic hyper-reduction.SAND2019-5878 M

Blonigan, Patrick↗

Microreactor Automated Control System Test Bed Digital Architecture for Real-Time, Hardware-in-the-Loop Simulation

This work describes progress made towards the development of a real-time hardware-in-the-loop (HIL) test bed for non-nuclear testing of microreactor control schemes and failure modes. Non-nuclear testing is a crucial step in developing robust control algorithms for managing microreactor dynamics. The creation of an HIL simulation harnesses the realistic dynamics of physical analogue systems while additionally considering the challenges of variable communication delay. This collaborative effort between Oak Ridge National Laboratory and Idaho National Laboratory has resulted in a LabVIEW-based gRPC communication protocol which couples a TRANSFORM Modelica simulation of nuclear components to the ViBRANT physical hardware for realistic feedback and visual representation of control action in real time. A modular python client structure is developed to manage FMU-based Modelica simulation and real-time gRPC communication. HIL testing suggests that the modeled reactor with natural convection molten salt loop coolant configuration responds well to PID control of drum positioning for modulation of reactor core power, however, future efforts will be made to explore the added thermal inertial delay of system level control and downstream demand changes. Development of this platform with a generalized methodology provides a foundation for exploring a variety of reactor configurations and failure modes in rapid order to provide insight into the most effective avenues of study for further research and development.

McConnell, Jono [ORNL] (ORCID:0000000238984741)↗

Microreactor Automated Control System - Digital Twin Models and Advanced Control Systems Updates

Automation of control systems is expected to be important in the economic and safe operation of microreactors. Therefore, there is a need to develop and demonstrate automated control for microreactors, along with the development of testbeds for this purpose. This report provides updates on the status of a nonnuclear microreactor automated control system (MACS)—a real-time, hardware-in-the-loop testbed for non-nuclear testing of microreactor control system automation. A real-time hardware-in-the-loop testbed incorporates the realistic dynamics of physical systems into control system development and testing. The collaborative effort between Oak Ridge National Laboratory (ORNL) and Idaho National Laboratory (INL) resulted in the development of a prototypic microreactor plant-level digital twin that includes the reactor and a balance of plant system. Advanced control strategies were incorporated to demonstrate testing of control automation solutions. The gRPC communication protocol, which was implemented in the hardware-in-the-loop testbed by INL, was coupled to a digital twin model developed using the TRANsient Simulation Framework of Reconfigurable Models (TRANSFORM) library in Modelica. This digital twin simulation was tested with the ViBRANT hardware for realistic feedback and visual representation of control action in real time. A modular Python client structure was developed to manage functional mock-up unit-based simulation and real-time gRPC communication. Hardware-in-the-loop testing indicated that the modeled reactor—a natural-convection, molten-salt coolant loop configuration—responds well to control of drum positioning for modulation of reactor core power, as well as system-level control and downstream demand changes. Ongoing research is focused on integrating additional control algorithms that utilize data from newly included sensors within the MACS hardware testbed, as well as demonstrating and assessing the performance of the different automated control algorithms on multiple additional operational scenarios.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

HAL/S - The programming language for Shuttle

HAL/S is a higher order language and system, now operational, adopted by NASA for programming Space Shuttle on-board software. Program reliability is enhanced through language clarity and readability, modularity through program structure, and protection of code and data. Salient features of HAL/S include output orientation, automatic checking (with strictly enforced compiler rules), the availability of linear algebra, real-time control, a statement-level simulator, and compiler transferability (for applying HAL/S to additional object and host computers). The compiler is described briefly.

Martin, F. H.↗

Modular multipurpose panel support

Inexpensive set of modular semirigid box structures serve as multipurpose support for thin panels. Fixture holds several different, interchangeable, two-part, adjustable, contour-board subassemblies. Combining modules allows for accommodation to any length skin or panel.

Daun, M. R.↗

Modular bus and payload designs for the Shuttle era

The Space Shuttle, in combination with the Interim Upper Stage, will provide an economical means of placing payloads in geostationary orbit in the 1980s. This paper summarizes a recent study of multidiscipline applications payloads for Shuttle-IUS launch. Various concepts for modular 'bus' spacecraft structures and payload groupings based on equipment commonality are considered. Several bus and payload configurations, ranging from 160 to 1600 kg, are developed. A typical payload is described in some detail. The payload consists of equipment at 400 MHz for search and rescue and data collection, at 1.5 and 15 GHz for aeronautical, maritime, and land mobile communications, at 40 and 90 GHz for millimeter wave communications, and at other bands for meteorological radiometry, electromagnetic environment measurements, and interferometry.

Durrani, S. H.↗

F-15B/Flight Test Fixture 2: A Test Bed for Flight Research

NASA Dryden Flight Research Center has developed a second-generation flight test fixture for use as a generic test bed for aerodynamic and fluid mechanics research. The Flight Test Fixture 2 (FTF-2) is a low-aspect-ratio vertical fin-like shape that is mounted on the centerline of the F-I5B lower fuselage. The fixture is designed for flight research at Mach numbers to a maximum of 2.0. The FTF-2 is a composite structure with a modular configuration and removable components for functional flexibility. This report documents the flow environment of the fixture, such as surface pressure distributions and boundary-layer profiles, throughout a matrix of conditions within the F-15B/FTF-2 flight envelope. Environmental conditions within the fixture are presented to assist in the design and testing of future avionics and instrumentation. The intent of this document is to serve as a user's guide and assist in the development of future flight experiments that use the FTF-2 as a test bed. Additional information enclosed in the appendices has been included to assist with more detailed analyses, if required.

David M Richwine↗

Development of Improved Models, Stochasticity, and Frameworks for the MIT Extensible Air Network Simulation

MEANS, the MIT Extensible Air Network Simulation, was created in February of 2001, and has been developed with support from NASA Ames since August of 2001. MEANS is a simulation tool which is designed to maximize fidelity without requiring data of such a low level as to preclude easy examination of alternative scenarios. To this end, MEANS is structured in a modular fashion to allow more detailed components to be brought in when desired, and left out when they would only be an impediment. Traditionally, one of the difficulties with high-fidelity models is that they require a level of detail in their data that is difficult to obtain. For analysis of past scenarios, the required data may not have been collected, or may be considered proprietary and thus difficult for independent researchers to obtain. For hypothetical scenarios, generation of the data is sufficiently difficult to be a task in and of itself. Often, simulations designed by a researcher will model exactly one element of the problem well and in detail, while assuming away other parts of the problem which are not of interest or for which data is not available. While these models are useful for working with the task at hand, they are very often not applicable to future problems. The MEAN Simulation attempts to address these problems by using a modular design which provides components of varying fidelity for each aspect of the simulation. This allows for the most accurate model for which data is available to be used. It also provides for easy analysis of sensitivity to data accuracy. This can be particularly useful in the case where accurate data is available for some subset of the situations that are to be considered. Furthermore, the ability to use the same model while examining effects on different parts of a system reduces the time spent learning the simulation, and provides for easier comparisons between changes to different parts of the system.

Clarke, John-Paul↗