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At least 235 records · Page 13

Use of non-spherical gravity harmonics for relative motion GN/C (category 2)

Detailed analysis of the Automatic Rendezvous and Capture problem indicate a need for three different regions of mathematical description for the GN&C algorithms: (1) multi-vehicle orbital mechanics to the rendezvous interface point, i.e., within 100 nm; (2) relative motion solutions (such as Clohessy-Wiltshire type) from the far-field to the near-field interface, i.e., within 1 nm and; (3) close proximity motion - the near-field motion where the relative differences in the gravitational and orbit inertial accelerations can be neglected from the equations of motion. Limit boundaries to these regions can be precisely defined by further analysis and will be functions of the tracking measurement accuracies and the computer resources available for the solution of the algorithms. This paper analyzes the relative motion in Regions 2 and 3 above and present the derivation and discussion of the general case of non-spherical gravitational perturbed relative motion. Mathematical deviations from the numerically integrated spherical gravity case and solutions from the Clohessy-Wiltshire equations are presented in the analysis. Based upon this preliminary analysis, it is recommended that further efforts be used to assess the relative position and velocity differences in Region 2 due to non-spherical gravity harmonics and that viable GN&C algorithms be developed to include these gravity perturbations (especially the effects of the first gravity harmonic, J2).

Henderson, David M.↗

Boron adsorption in clay minerals: Implications for martian groundwater chemistry and boron on Mars

Boron has been detected on Mars in calcium-sulfate veins found within clay mineral rich rocks on Mars by the Mars Science Laboratory (MSL) Curiosity rover using Laser Induced Breakdown Spectroscopy (LIBS) analysis. Borates play a vital role in stabilizing ribose on Earth and has been suggested as a key requirement for life. Borate ions readily adsorb to phyllosilicate clay minerals. The discovery of boron on Mars in proximity to phyllosilicate-bearing bedrock may have strong implications for potential past prebiotic conditions on Mars. In this study we generated a suite of clay minerals with adsorbed borate, including both typical terrestrial clay minerals (montmorillonite) and Mars-analog clay minerals (nontronite, saponite, griffithite), to understand controls on borate adsorption and to analyze with LIBS to compare with MSL data. Clay minerals were subjected to mineralogical and chemical analysis before and after adsorption. Adsorption analysis revealed that the Mars analog clay minerals adsorbed less boron than terrestrial counterparts, but within comparable amounts to those detected on Mars and in meteorites. Post-adsorption analysis by X-ray diffraction (XRD) revealed slight changes in the interlayer spacing of many of the clay minerals. Based on the adsorption analysis of the Mars-analog clay minerals, phyllosilicate-bearing bedrock in Gale crater may contain up to 90-110 ppm B. A series of borate-enriched samples were created for analysis of LIBS spectra from ChemCam on the Curiosity rover and SuperCam on the Perseverance Rover. Finally, the results of this study may provide insight into martian groundwater geochemistry processes and the mobility of a key molecule connected with life.

58 GEOSCIENCES↗

Model-based Hierarchical Reinforcement Learning for Improved Physical Security Design: A Prototype

Prior work in FY24 developed an adversarial AI agent aid in path analysis of physical protection systems. This agent, trained using a model-based reinforcement learning algorithm, was able to successfully learn the most vulnerable path in facilities. It was able to extend the current state of practice for physical protection design by exhibiting dynamic behavior based on current environmental conditions. Whereas PathTrace largely performs a static, graph-based analysis, the AI agent was able to make decisions based on relative position in the facility, current conditions (was the adversarial agnet discovered?), and proximity to secondary targets. The agent demonstrated some novel capabilities, but had limitations that need to be resolved before it can be used for production purposes. For example, the adversarial agent generalizes poorly and takes a relatively long time to train. Nonetheless, there is still considerable promise for developing the adversarial agent further in order to explore even richer, more dynamic behaviors (e.g., adversary motivations, environmental debris, and more). This work considers a complementary idea; development of a planning agent. The planning agent is envisioned as an auto-complete-like tool that can help accelerate security system design by human experts. The agent would respect existing barriers and sensors placed by a human expert while offering cost-effective suggestions (i.e., implicitly balancing effectiveness with cost) to improve the design. The goal is for this agent to be part of an expert’s toolbox, not to totally upend the current state-of-practice, or to displace human experts. The ultimate goal would be concurrent training of both the adversarial and planning agent together, to learn entirely through self-play. This would represent an entirely new way of performing system deign. We selected a hierarchical, model-based reinforcement learning algorithm to serve as the planning agent. This is an extension of concepts used in the prior FY24 adversarial agent work. There, we had a single agent acting an environment. Here, we have two different sub-agents (policies), working together, to form a complete agent. There is a manager policy, which can select abstract goals on slower time scales, and a worker, which performs primitive actions to reach goals selected by the manager. It is worth noting that this class of algorithm is challenging to work with. From our understanding, our work is one of the first successful uses of model-based reinforcement learning (MBRL) in nuclear energy1 , and likely the first hierarchical model-based reinforcement learning application in nuclear energy. Further, this work is one of the first known attempts to apply AI to perform a design tasks in nuclear energy. Consequently, there were significant implementation challenges and the bulk of the work was focused on successful implementation and algorithm design. The results presented here are very low technology readiness level as a consequence of the lack of related literature, but still represent a significant step forward in the pursuit of applied AI for design.

42 ENGINEERING↗

Knowledge-based vision for space station object motion detection, recognition, and tracking

Computer vision, especially color image analysis and understanding, has much to offer in the area of the automation of Space Station tasks such as construction, satellite servicing, rendezvous and proximity operations, inspection, experiment monitoring, data management and training. Knowledge-based techniques improve the performance of vision algorithms for unstructured environments because of their ability to deal with imprecise a priori information or inaccurately estimated feature data and still produce useful results. Conventional techniques using statistical and purely model-based approaches lack flexibility in dealing with the variabilities anticipated in the unstructured viewing environment of space. Algorithms developed under NASA sponsorship for Space Station applications to demonstrate the value of a hypothesized architecture for a Video Image Processor (VIP) are presented. Approaches to the enhancement of the performance of these algorithms with knowledge-based techniques and the potential for deployment of highly-parallel multi-processor systems for these algorithms are discussed.

Symosek, P.↗

An active role of the amyloplasts and nuclei of root statocytes in graviperception

Three main phases are discerned in the gravitropic reaction: perception of a gravitational stimulus, its transduction, and fixation of the reaction resulting in bending of an organ. According to the starch-statolith hypothesis of Nemec and Haberlandt, amyloplasts in the structurally and functionally specialized graviperceptive cells (statocytes) sediment in the direction of a gravitational vector in the distal part of a cell while a nucleus is in the proximal one. If amyloplasts appear to act as gravity sensors, the receptors, which interact with sedimented amyloplasts, and next signaling are still unclear. An analysis of the structural-functional organization of cells in different root cap layers of such higher plants as pea, Arabidopsis thaliana, and Brassica rapa grown under 1 g, on the clinostats, and in microgravity, allows us to support the hypothesis that amyloplasts function as statoliths in statocytes, but they may not be only the passive statolithic mass. We propose that amyloplasts fulfill a more complex function by interacting with a receptor, which is a nucleus, in transduction of some signal to it. Gravity-induced statolith movement in certain order leads to a new functional connection between gravity susceptors--amyloplasts and a receptor--a nucleus receiving some signal presumedly of a mechanical or biochemical nature from the amyloplasts. During gravitropism, sugar signaling could induce expression of genes encoding auxin transport proteins in a nucleus giving the nucleus an intermediate role in signal trunsduction following perception. c 2001 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

short duration↗

Practical Methodology for the Inclusion of Nonlinear Slosh Damping in the Stability Analysis of Liquid-Propelled Space Vehicles

One of the challenges of developing flight control systems for liquid-propelled space vehicles is ensuring stability and performance in the presence of parasitic minimally damped slosh dynamics in the liquid propellants. This can be especially difficult when the fundamental frequencies of the slosh motions are in proximity to the frequency used for vehicle control. The challenge is partially alleviated since the energy dissipation and effective damping in the slosh modes increases with amplitude. However, traditional launch vehicle control design methodology is performed with linearized systems using a fixed slosh damping corresponding to a slosh motion amplitude based on heritage values. This papers presents a method for performing the control design and analysis using damping at slosh amplitudes chosen based on the resulting limit cycle amplitude of the vehicle thrust vector system due to a control-slosh interaction under degraded phase and gain margin conditions.

Ottander, John A.↗

Practical Methodology for the Inclusion of Nonlinear Slosh Damping in the Stability Analysis of Liquid-propelled Space Vehicles

One of the challenges of developing flight control systems for liquid-propelled space vehicles is ensuring stability and performance in the presence of parasitic minimally damped slosh dynamics in the liquid propellants. This can be especially difficult when the fundamental frequencies of the slosh motions are in proximity to the frequency used for vehicle control. The challenge is partially alleviated since the energy dissipation and effective damping in the slosh modes increases with amplitude. However, traditional launch vehicle control design methodology is performed with linearized systems using a fixed slosh damping corresponding to a slosh motion amplitude based on heritage values. This papers presents a method for performing the control design and analysis using damping at slosh amplitudes chosen based on the resulting limit cycle amplitude of the vehicle thrust vector system due to a control-slosh interaction under degraded phase and gain margin conditions.

Ottander, John A.↗

Time Dependence of Collision Probabilities During Satellite Conjunctions

The NASA Conjunction Assessment Risk Analysis (CARA) team has recently implemented updated software to calculate the probability of collision (P (sub c)) for Earth-orbiting satellites. The algorithm can employ complex dynamical models for orbital motion, and account for the effects of non-linear trajectories as well as both position and velocity uncertainties. This “3D P (sub c)” method entails computing a 3-dimensional numerical integral for each estimated probability. Our analysis indicates that the 3D method provides several new insights over the traditional “2D P (sub c)” method, even when approximating the orbital motion using the relatively simple Keplerian two-body dynamical model. First, the formulation provides the means to estimate variations in the time derivative of the collision probability, or the probability rate, R (sub c). For close-proximity satellites, such as those orbiting in formations or clusters, R (sub c) variations can show multiple peaks that repeat or blend with one another, providing insight into the ongoing temporal distribution of risk. For single, isolated conjunctions, R (sub c) analysis provides the means to identify and bound the times of peak collision risk. Additionally, analysis of multiple actual archived conjunctions demonstrates that the commonly used “2D P (sub c)” approximation can occasionally provide inaccurate estimates. These include cases in which the 2D method yields negligibly small probabilities (e.g., P (sub c)) is greater than 10 (sup -10)), but the 3D estimates are sufficiently large to prompt increased monitoring or collision mitigation (e.g., P (sub c) is greater than or equal to 10 (sup -5)). Finally, the archive analysis indicates that a relatively efficient calculation can be used to identify which conjunctions will have negligibly small probabilities. This small-P (sub c) screening test can significantly speed the overall risk analysis computation for large numbers of conjunctions.

Hall, Doyle T.↗

Risk Analysis of a Hydrogen Generation Facility near a Nuclear Power Plant

Nuclear power plants (NPPs) are considering flexible plant operations to take advantage of excess thermal and electrical energy. One option for NPPs is to pursue hydrogen production through high temperature electrolysis as an alternate revenue stream to remain economically viable. The intent of this study is to investigate the risk of a hydrogen production facility in close proximity to an NPP. A 100 MW, 500 MW, and 1,000 MW facility are evaluated herein. Previous analyses have evaluated preliminary designs of a hydrogen production facility in a conservative manner to determine if it is feasible to co-locate the facility within 1 km of an NPP. This analysis specifically evaluates the risk components of different hydrogen production facility designs, including the likelihood of a leak within the system and the associated consequence to critical NPP targets. This analysis shows that although the likelihood of a leak in an HTEF is not negligible, the consequence to critical NPP targets is not expected to lead to a failure given adequate distance from the plant.

08 HYDROGEN↗

Development and verification of local/global analysis techniques for laminated composites

Analysis and design methods for laminated composite materials have been the subject of considerable research over the past 20 years, and are currently well developed. In performing the detailed three-dimensional analyses which are often required in proximity to discontinuities, however, analysts often encounter difficulties due to large models. Even with the current availability of powerful computers, models which are too large to run, either from a resource or time standpoint, are often required. There are several approaches which can permit such analyses, including substructuring, use of superelements or transition elements, and the global/local approach. This effort is based on the so-called zoom technique to global/local analysis, where a global analysis is run, with the results of that analysis applied to a smaller region as boundary conditions, in as many iterations as is required to attain an analysis of the desired region. Before beginning the global/local analyses, it was necessary to evaluate the accuracy of the three-dimensional elements currently implemented in the Computational Structural Mechanics (CSM) Testbed. It was also desired to install, using the Experimental Element Capability, a number of displacement formulation elements which have well known behavior when used for analysis of laminated composites.

Griffin, O. Hayden, Jr.↗

Spectral Energy Distribution and Bolometric Luminosity of the Cool Brown Dwarf Gliese 229B

Infrared broadband photometry of the cool brown dwarf Gliese 229B extending in wavelength from 0.8 to 10.5 micron is reported. These results are derived from both new data and reanalyzed, previously published data. Existing spectral data reported have been rereduced and recalibrated. The close proximity of the bright Gliese 229A to the dim Gliese 229B required the use of special techniques for the observations and also for the data analysis. We describe these procedures in detail. The observed luminosity between 0.8 and 10.5 micron is (4.9 +/- 0.6) x 10(exp -6) solar luminosity. The observed spectral energy distribution is in overall agreement with a dust-free model spectrum by Tsuji et al. for T(eff) approx. equal to 900 K. If this model is used to derive the bolometric correction, the best estimate of the bolometric luminosity is 6.4 x 10(exp -6) solar luminosity and 50% of this luminosity ties between 1 and 2.5 microns. Our best estimate of the effective temperature is 900 K. From the observed near-infrared spectrum and the spectral energy distribution, the brightness temperatures (T(sub B) are estimated. The highest, T(sub B) = 1640 K, is seen at the peak of the J band spectrum, while the lowest, T(sub B) is less than or equal to 600 K, is at 3.4 microns, which corresponds to the location of the fundamental methane band.

Matthews, K.↗

Trajectory Design Employing Convex Optimization for Landing on Irregularly Shaped Asteroids

Mission proposals that land on asteroids are becoming popular. However, in order to have a successful mission the spacecraft must reliably and softly land at the intended landing site. The problem under investigation is how to design a fuel-optimal powered descent trajectory that can be quickly computed on- board the spacecraft, without interaction from ground control. An optimal trajectory designed immediately prior to the descent burn has many advantages. These advantages include the ability to use the actual vehicle starting state as the initial condition in the trajectory design and the ease of updating the landing target site if the original landing site is no longer viable. For long trajectories, the trajectory can be updated periodically by a redesign of the optimal trajectory based on current vehicle conditions to improve the guidance performance. One of the key drivers for being completely autonomous is the infrequent and delayed communication between ground control and the vehicle. Challenges that arise from designing an asteroid powered descent trajectory include complicated nonlinear gravity fields, small rotating bodies and low thrust vehicles. There are two previous studies that form the background to the current investigation. The first set looked in-depth at applying convex optimization to a powered descent trajectory on Mars with promising results.1, 2 This showed that the powered descent equations of motion can be relaxed and formed into a convex optimization problem and that the optimal solution of the relaxed problem is indeed a feasible solution to the original problem. This analysis used a constant gravity field. The second area applied a successive solution process to formulate a second order cone program that designs rendezvous and proximity operations trajectories.3, 4 These trajectories included a Newtonian gravity model. The equivalence of the solutions between the relaxed and the original problem is theoretically established. The proposed solution for designing the asteroid powered descent trajectory is to use convex optimization, a gravity model with higher fidelity than Newtonian, and an iterative solution process to design the fuel optimal trajectory. The solution to the convex optimization problem is the thrust profile, magnitude and direction, that will yield the minimum fuel trajectory for a soft landing at the target site, subject to various mission and operational constraints. The equations of motion are formulated in a rotating coordinate system and includes a high fidelity gravity model. The vehicle's thrust magnitude can vary between maximum and minimum bounds during the burn. Also, constraints are included to ensure that the vehicle does not run out of propellant, or go below the asteroid's surface, and any vehicle pointing requirements. The equations of motion are discretized and propagated with the trapezoidal rule in order to produce equality constraints for the optimization problem. These equality constraints allow the optimization algorithm to solve the entire problem, without including a propagator inside the optimization algorithm.

Pinson, Robin M.↗

Investigation of Rotor-Airframe Interaction Noise Associated with Small-Scale Rotary-Wing Unmanned Aircraft Systems

In this study, hover acoustic measurements are taken on isolated rotor-airframe configurations representative of smallscale, rotary-wing unmanned aircraft systems (UAS). Each rotor-airframe configuration consists of two fixed-pitch blades powered by a brushless motor, with a simplified airframe geometry intended to represent a generic multicopter arm. In addition to acoustic measurements, CFD-based aeroacoustic predictions are implemented on a subset of the experimentally tested rotor-airframe configurations in an effort to better understand the noise content of the rotor-airframe systems. Favorable agreements are obtained between acoustic measurements and predictions, based on both time- and frequency-domain post-processing techniques. Results indicate that close proximity of airframe surfaces result in the generation of considerable tonal acoustic content in the form of harmonics of the rotor blade passage frequency (BPF). Analysis of the acoustic prediction data shows that the presence of the airframe surfaces can generate noise levels either comparable to or greater than the rotor blade surfaces under certain rotor tip clearance conditions. Analysis of the on-surface Ffowcs Williams and Hawkings (FW-H) source terms provide insight as to the predicted physical noise-generating mechanisms on the rotor and airframe surfaces.

Zawodny, Nikolas S.↗

FLIMJ: An open-source ImageJ toolkit for fluorescence lifetime image data analysis

In the field of fluorescence microscopy, there is continued demand for dynamic technologies that can exploit the complete information from every pixel of an image. One imaging technique with proven ability for yielding additional information from fluorescence imaging is Fluorescence Lifetime Imaging Microscopy (FLIM). FLIM allows for the measurement of how long a fluorophore stays in an excited energy state, and this measurement is affected by changes in its chemical microenvironment, such as proximity to other fluorophores, pH, and hydrophobic regions. This ability to provide information about the microenvironment has made FLIM a powerful tool for cellular imaging studies ranging from metabolic measurement to measuring distances between proteins. The increased use of FLIM has necessitated the development of computational tools for integrating FLIM analysis with image and data processing. To address this need, we have created FLIMJ, an ImageJ plugin and toolkit that allows for easy use and development of extensible image analysis workflows with FLIM data. Built on the FLIMLib decay curve fitting library and the ImageJ Ops framework, FLIMJ offers FLIM fitting routines with seamless integration with many other ImageJ components, and the ability to be extended to create complex FLIM analysis workflows. Building on ImageJ Ops also enables FLIMJ’s routines to be used with Jupyter notebooks and integrate naturally with science-friendly programming in, e.g., Python and Groovy. We show the extensibility of FLIMJ in two analysis scenarios: lifetime-based image segmentation and image colocalization. We also validate the fitting routines by comparing them against industry FLIM analysis standards.

47 OTHER INSTRUMENTATION↗

ASRM Multi-Port Igniter Flow Field Analysis

The Advanced Solid Rocket Motor (ASRM) program was initiated by NASA in response to the need for a new generation rocket motor capable of providing increased thrust levels over the existing Redesigned Solid Rocket Motor (RSRM) and thus augment the lifting capacity of the space shuttle orbiter. To achieve these higher thrust levels and improve motor reliability, advanced motor design concepts were employed. In the head end of the motor, for instance, the propellent cast has been changed from the conventional annular configuration to a 'multi-slot' configuration in order to increase the burn surface area and guarantee rapid motor ignition. In addition, the igniter itself has been redesigned and currently features 12 exhaust ports in order to channel hot igniter combustion gases into the circumferential propellent slots. Due to the close proximity of the igniter ports to the propellent surfaces, new concerns over possible propellent deformation and erosive burning have arisen. The following documents the effort undertaken using computational fluid dynamics to perform a flow field analysis in the top end of the ASRM motor to determine flow field properties necessary to permit a subsequent propellent fin deformation analysis due to pressure loading and an assessment of the extent of erosive burning.

Kania, Lee↗

Estimating Trabecular Bone Mechanical Properties From Non-Invasive Imaging

An important component in developing countermeasures for maintaining musculoskeletal integrity during long-term space flight is an effective and meaningful method of monitoring skeletal condition. Magnetic resonance imaging (MRI) is an attractive non-invasive approach because it avoids the exposure to radiation associated with X-ray based imaging and also provides measures related to bone microstructure rather than just density. The purpose of the research for the 1996 Summer Faculty Fellowship period was to extend the usefulness of the MRI data to estimate the mechanical properties of trabecular bone. The main mechanical properties of interest are the elastic modulus and ultimate strength. Correlations are being investigated between these and fractal analysis parameters, MRI relaxation times, apparent densities, and bone mineral densities. Bone specimens from both human and equine donors have been studied initially to ensure high-quality MR images. Specimens were prepared and scanned from human proximal tibia bones as well as the equine distal radius. The quality of the images from the human bone appeared compromised due to freezing artifact, so only equine bone was included in subsequent procedures since these specimens could be acquired and imaged fresh before being frozen. MRI scans were made spanning a 3.6 cm length on each of 5 equine distal radius specimens. The images were then sent to Dr. Raj Acharya of the State University of New York at Buffalo for fractal analysis. Each piece was cut into 3 slabs approximately 1.2 cm thick and high-resolution contact radiographs were made to provide images for comparing fractal analysis with MR images. Dual energy X-ray absorptiometry (DEXA) scans were also made of each slab for subsequent bone mineral density determination. Slabs were cut into cubes for mechanical using a slow-speed diamond blade wafering saw (Buehler Isomet). The dimensions and wet weights of each cube specimen were measured and recorded. Wet weights were also recorded. Each specimen was labeled and marked to denote anatomic orientations, i.e. superior/inferior (S/I), media/lateral (M/L), and anterior/posterior (A/P). The actual locations of each cube cut were documented and images distributed to define ROI locations for other analyses (to Raj Acharya for fractal analysis, to Jon Richardson at Baylor College of Medicine for DEXA, and to Chen Lin at Baylor College of Medicine for T2* MRI analysis). Quasistatic mechanical testing consisted of compressive loading in all three mutually perpendicular anatomic directions. Cyclic loading was applied for 10 cycles to precondition the specimen and results calculated for the eleventh. For one of three directions tested on each specimen, the 10 cycles were followed with loading to failure. Testing is currently proceeding and once completed the results will be correlated with data from the other analyses. One of the main points of interest is the relationship between fractal dimension and mechanical properties. Throughout preparation and testing all specimens were maintained hydrated with physiological saline and stored frozen when not being used.

Hogan, Harry A.↗

An experimental investigation of the transverse pressure gradient associated with curved free jets in the proximity of solid boundaries

An experiment has been performed to determine the mass flow and thrust augmentation associated with the flow of a two-dimensional incompressible jet issued parallel to an adjacent offset wall. A theoretical analysis based on a linear static pressure profile across the jet exit reveals that thrust augmentation is possible only for a wall offset of less than one half of the nozzle exit width. Mass flow augmentation measurements are in reasonable agreement with the proposed theory. Augmented mass flow of over three percent is observed directly, while thrust augmentation in excess of three percent is predicted when measured values of the pressure difference across the exiting jet are supplied to the theory. Measurements of the separation cavity length are presented which complement those given by previous investigators by extending the range to small values of the ratio of the wall offset to the slot height. For large offset ratios it is found that the cavity length approaches 1.25 times the wall offset distance. The initial radius of curvature of the jet centerline is found to be substantially greater than the mean radius of curvature over the cavity length, thereby indicating that the trajectory of the jet does not follow the arc of a circle as has been previously assumed.

Lund, T. S.↗

Mutation of active site glutamate in serine hydroxymethyltransferase allows trapping a reactive intermediate: a combined neutron and X-ray crystallography study

Serine hydroxymethyltransferase (SHMT) is a pyridoxal-5′-phosphate (PLP) dependent enzyme that catalyzes a chemical transformation essential for the one-carbon (1C) metabolism. SHMT reversibly converts L-Ser into Gly and transfers a 1C unit to tetrahydrofolate (THF) to give 5,10-methylene-THF (5,10-MTHF). 5,10-MTHF, a 1C-unit donor, plays a crucial role in the downstream biomolecular syntheses required for the cell homeostasis and proliferation. SHMT is a prominent target for the drug discovery to battle bacterial and parasitic infections, and to treat various types of cancer. SHMT-catalyzed chemistry is governed by the general acid-base catalysis. Knowledge of the catalytic mechanism can aid drug design but can only be achieved when the atomic details of each reaction step are mapped, including accurate determination of hydrogen atom positions. Here we utilized the inactive E53Q mutant of Thermus thermophilus ( Tth ) SHMT to directly determine protonation states with room-temperature neutron crystallography and to capture a reactive intermediate containing the PLP-L-Ser external aldimine and THF in the enzyme active site. We observed protonation of the Schiff base nitrogen (N SB ) in the PLP internal aldimine but no change in the protonation states of other ionizable PLP groups and active site residues compared to wild-type Tth SHMT. X-ray structural analysis of the ternary intermediate complex E53Q-Ser-THF that eluded previous structural characterization shows the strategic positioning of the E53Q side chain in close proximity to the external aldimine and THF and reinforces the proposed role for E53 as the driver of proton transfer events along the reaction pathway.

Drago, Victoria N. [Oak Ridge National Laboratory ↗