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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 199 records · Page 11

De novo design of pH-responsive self-assembling helical protein filaments

Abstract Biological evolution has led to precise and dynamic nanostructures that reconfigure in response to pH and other environmental conditions. However, designing micrometre-scale protein nanostructures that are environmentally responsive remains a challenge. Here we describe the de novo design of pH-responsive protein filaments built from subunits containing six or nine buried histidine residues that assemble into micrometre-scale, well-ordered fibres at neutral pH. The cryogenic electron microscopy structure of an optimized design is nearly identical to the computational design model for both the subunit internal geometry and the subunit packing into the fibre. Electron, fluorescent and atomic force microscopy characterization reveal a sharp and reversible transition from assembled to disassembled fibres over 0.3 pH units, and rapid fibre disassembly in less than 1 s following a drop in pH. The midpoint of the transition can be tuned by modulating buried histidine-containing hydrogen bond networks. Computational protein design thus provides a route to creating unbound nanomaterials that rapidly respond to small pH changes.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

The CNK–HYP scaffolding complex promotes RAF activation by enhancing KSR–MEK interaction

The RAS–MAPK pathway regulates cell proliferation, differentiation and survival, and its dysregulation is associated with cancer development. The pathway minimally comprises the small GTPase RAS and the kinases RAF, MEK and ERK. Activation of RAF by RAS is notoriously intricate and remains only partially understood. There are three RAF isoforms in mammals (ARAF, BRAF and CRAF) and two related pseudokinases (KSR1 and KSR2). RAS-mediated activation of RAF depends on an allosteric mechanism driven by the dimerization of its kinase domain. Recent work on human RAFs showed that MEK binding to KSR1 promotes KSR1–BRAF heterodimerization, which leads to the phosphorylation of free MEK molecules by BRAF. Similar findings were made with the single Drosophila RAF homolog. Here we show that the fly scaffold proteins CNK and HYP stabilize the KSR–MEK interaction, which in turn enhances RAF–KSR heterodimerization and RAF activation. The cryogenic electron microscopy structure of the minimal KSR–MEK–CNK–HYP complex reveals a ring-like arrangement of the CNK–HYP complex allowing CNK to simultaneously engage KSR and MEK, thus stabilizing the binary interaction. Together, these results illuminate how CNK contributes to RAF activation by stimulating the allosteric function of KSR and highlight the diversity of mechanisms impacting RAF dimerization as well as the regulatory potential of the KSR–MEK interaction.

59 BASIC BIOLOGICAL SCIENCES↗

Atomic view of photosynthetic metabolite permeability pathways and confinement in synthetic carboxysome shells

Carboxysomes are protein microcompartments found in cyanobacteria, whose shell encapsulates rubisco at the heart of carbon fixation in the Calvin cycle. Carboxysomes are thought to locally concentrate CO 2 in the shell interior to improve rubisco efficiency through selective metabolite permeability, creating a concentrated catalytic center. However, permeability coefficients have not previously been determined for these gases, or for Calvin-cycle intermediates such as bicarbonate (HCO$^–_3$), 3-phosphoglycerate, or ribulose-1,5-bisphosphate. Starting from a high-resolution cryogenic electron microscopy structure of a synthetic β-carboxysome shell, we perform unbiased all-atom molecular dynamics to track metabolite permeability across the shell. The synthetic carboxysome shell structure, lacking the bacterial microcompartment trimer proteins and encapsulation peptides, is found to have similar permeability coefficients for multiple metabolites, and is not selectively permeable to HCO$^–_3$ relative to CO 2 . To resolve how these comparable permeabilities can be reconciled with the clear role of the carboxysome in the CO 2 -concentrating mechanism in cyanobacteria, complementary atomic-resolution Brownian Dynamics simulations estimate the mean first passage time for CO 2 assimilation in a crowded model carboxysome. Despite a relatively high CO 2 permeability of approximately 10 -2 cm/s across the carboxysome shell, the shell proteins reflect enough CO 2 back toward rubisco that 2,650 CO 2 molecules can be fixed by rubisco for every 1 CO 2 molecule that escapes under typical conditions. The permeabilities determined from all-atom molecular simulation are key inputs into flux modeling, and the insight gained into carbon fixation can facilitate the engineering of carboxysomes and other bacterial microcompartments for multiple applications.

59 BASIC BIOLOGICAL SCIENCES↗

Design and structural basis of selective 1,4-dihydropyridine inhibitors of the calcium-activated potassium channel K Ca 3.1

The 1,4-dihydropyridines, drugs with well-established bioavailability and toxicity profiles, have proven efficacy in treating human hypertension, peripheral vascular disorders, and coronary artery disease. Every 1,4-dihydropyridine in clinical use blocks L-type voltage-gated calcium channels. We now report our development, using selective optimization of a side activity (SOSA), of a class of 1,4-dihydropyridines that selectively and potently inhibit the intermediate-conductance calcium-activated K + channel K Ca 3.1, a validated therapeutic target for diseases affecting many organ systems. One of these 1,4-dihydropyridines, DHP-103, blocked K Ca 3.1 with an IC 50 of 6 nM and exhibited exquisite selectivity over calcium channels and a panel of >100 additional molecular targets. Using high-resolution structure determination by cryogenic electron microscopy together with mutagenesis and electrophysiology, we delineated the drug binding pocket for DHP-103 within the water-filled central cavity of the K Ca 3.1 channel pore, where bound drug directly impedes ion permeation. DHP-103 inhibited gain-of-function mutant K Ca 3.1 channels that cause hereditary xerocytosis, suggesting its potential use as a therapeutic for this hemolytic anemia. In a rat model of acute ischemic stroke, the second leading cause of death worldwide, DHP-103 administered 12 h postischemic insult in proof-of-concept studies reduced infarct volume, improved balance beam performance (measure of proprioception) and decreased numbers of activated microglia in infarcted areas. K Ca 3.1-selective 1,4-dihydropyridines hold promise for the many diseases for which K Ca 3.1 has been experimentally confirmed as a therapeutic target.

Ong, Seow Theng [Lee Kong Chian School of Medicine↗

Design alternatives for the Shuttle Infrared Telescope Facility

The paper discusses the Shuttle Infrared Telescope Facility (SIRTF), a versatile astronomical telescope that can accomodate photometric, spectroscopic, and polarimetric measurements. It is expected to be 100 to 1000 times more sensitive than any existing infrared telescope; detailed designs of cooled IR telescopes were made for the Infrared Astronomical Satellite and the Small Helium Cooled Infrared Telescope for Spacelab 2. Rocket tests verified the capability of using superfluid helium as a cryogen in zero gravity. Constraints on funds for Shuttle payloads require an evolutionary approach to the development of the full potential of SIRTF, necessitating consideration of design alternatives involving the optical configuration, the cryogen, the mechanical structure, and size of SIRTF.

Witteborn, F. C.↗

Aerospace vehicle

A dual structure aerospace vehicle is described which has an aeroshell structure and an internally disposed separable and reusable integral tank/thrust structure. The tank/thrust structure is inuslated for cryogenic fuels and the cavity within aeroshell is insulated from the tank/thrust structure. An internal support ring within the cavity serves as an attachment for lugs on the tank/thrust structure via double hinges. The aft end of tank/thrust structure is provided with rocket engines and exit nozzles with a trunnion supporting the tank/thrust structure within the aeroshell.

Jackson, L. R.↗

The design of a precision pointing contol system for the Space Infrared Telescope Facility

The design and performance simulation of the Space Infrared Telescope Facility pointing and control system are described. The system design is constrained by such considerations as image stability, maneuver capability, cryogen lifetime, and structural flexibility. The primary design driver is the requirement to rapidly execute small angle reorientations of the telescope's optical axis. The flexibility of the spacecraft was simulated using a simplified multibody model which provided an approximation of the primary bending modes. The rather conventional attitude control system design of PID type with a 0.5 Hz bandwidth was enhanced to meet the small angle maneuver requirements by the use of three techniques: (1) a feedforward loop to improve the time response; (2) torque shaping to minimize structural excitation; and (3) active image stabilization to remove residual pointing error and jitter. Simulation results indicate that the system has the desired performance and can be designed using techniques and hardware well within the state-of-the-art.

Sridhar, B.↗

Analysis and test of low profile aluminum aerospace tank dome

In order to increase the structural performance of cryogenic tanks, the aerospace industry is beginning to employ low-profile bulkheads in new generation launch vehicle designs. This report details the analysis and test of one such dome made from 2219 aluminum. Such domes have two potential failure modes under internal pressure, general tensile failure and hoop compression buckling (in regions near the equator). The test determined the buckling load and ultimate tensile load of the hardware and showed that both compared well with the analysis predictions. This effort was conducted under the auspices of NASA and the General Dynamics Cryogenic Tank Technology Program (CTTP).

Ahmed, R.↗

X-33 Program Overview

An X-33 program overview is presented in viewgraph form. The objective of the research was to build and test a 50% scale prototype of an operational Reusable Launch Vehicle in a realistic flight environment. Technologies to be demonstrated include reusable cryogenic tankage, composite structures, durable thermal protection systems, advanced avionics, reliable propulsion systems, and aircraft-like operations.

Naftel, J. Chris↗

Automated Fiber Placement of PEEK/IM7 Composites with Film Interleaf Layers

The incorporation of thin discrete layers of resin between plies (interleafing) has been shown to improve fatigue and impact properties of structural composite materials. Furthermore, interleafing could be used to increase the barrier properties of composites used as structural materials for cryogenic propellant storage. In this work, robotic heated-head tape placement of PEEK/IM7 composites containing a PEEK polymer film interleaf was investigated. These experiments were carried out at the NASA Langley Research Center automated fiber placement facility. Using the robotic equipment, an optimal fabrication process was developed for the composite without the interleaf. Preliminary interleaf processing trials indicated that a two-stage process was necessary; the film had to be tacked to the partially-placed laminate then fully melted in a separate operation. Screening experiments determined the relative influence of the various robotic process variables on the peel strength of the film-composite interface. Optimization studies were performed in which peel specimens were fabricated at various compaction loads and roller temperatures at each of three film melt processing rates. The resulting data were fitted with quadratic response surfaces. Additional specimens were fabricated at placement parameters predicted by the response surface models to yield high peel strength in an attempt to gage the accuracy of the predicted response and assess the repeatability of the process. The overall results indicate that quality PEEK/lM7 laminates having film interleaves can be successfully and repeatability fabricated by heated head automated fiber placement.

Hulcher, A. Bruce↗

Simultaneous Temperature and Strain Sensing for Cryogenic Applications Using Dual-Wavelength Fiber Bragg Gratings

A new technique has been developed for sensing both temperature and strain simultaneously by using dual-wavelength fiber-optic Bragg gratings. Two Bragg gratings with different wavelengths were inscribed at the same location in an optical fiber to form a sensor. By measuring the wavelength shifts that resulted from the fiber being subjected to different temperatures and strains, the wavelength-dependent thermo-optic coefficients and photoelastic coefficients of the fiber were determined. This enables the simultaneous measurement of temperature and strain. In this study, measurements were made over the temperature range from room temperature down to about 10 K, addressing much of the low temperature range of cryogenic tanks. A structure transition of the optical fiber during the temperature change was found. This transition caused splitting of the waveforms characterizing the Bragg gratings, and the determination of wavelength shifts was consequently complicated. The effectiveness and sensitivities of these measurements in different temperature ranges are also discussed.

Wu, Meng-Chou↗

Using Dual-wavelength Fiber Bragg Gratings for Temperature and Strain Sensing at Cryogenic Temperature

By using dual-wavelength fiber-optic Bragg gratings, a new technique has been developed for sensing both temperature and strain simultaneously in cryogenic temperature range. Two Bragg gratings with different wavelengths were inscribed at the same location in an optical fiber to form a dual-wavelength sensor. By measuring the wavelength shifts that resulted from the fiber being subjected to different temperatures and strains, the wavelength-dependent thermo-optic coefficients and photoelastic coefficients of the fiber were determined. These coefficients were used to construct the elements of the K matrix, which enables to determine inversely the strain and temperature changes by measuring the wavelength shifts of the dual-wavelength Bragg grating. In this study, measurements were made over the temperature range from room temperature down to about 10 K, addressing much of the low temperature range of cryogenic tanks. A structure transition of the optical fiber during the temperature change was found from about 70 K to 140 K. This transition caused splitting of the waveforms characterizing the Bragg gratings, and the determination of wavelength shifts was consequently complicated. Several alternatives are proposed to resolve this problem. The effectiveness and sensitivities of these measurements in different temperature ranges are discussed. The separation of two wavelengths for the dual-wavelength Bragg grating has been widened to increase the sensitivities of measurement; however, this separation can still be covered in the scanning range from single scanning laser.

Wu, Meng-Chou↗

Methods and Piezoelectric Imbedded Sensors for Damage Detection in Composite Plates Under Ambient and Cryogenic Conditions

New methods for structural health monitoring are being assessed, especially in high-performance, extreme environment, safety-critical applications. One such application is for composite cryogenic fuel tanks. The work presented here attempts to characterize and investigate the feasibility of using imbedded piezoelectric sensors to detect cracks and delaminations under cryogenic and ambient conditions. A variety of damage detection methods and different Sensors are employed in the different composite plate samples to aid in determining an optimal algorithm, sensor placement strategy, and type of imbedded sensor to use. Variations of frequency, impedance measurements, and pulse echoing techniques of the sensors are employed and compared. Statistical and analytic techniques are then used to determine which method is most desirable for a specific type of damage. These results are furthermore compared with previous work using externally mounted sensors. Results and optimized methods from this work can then be incorporated into a larger composite structure to validate and assess its structural health. This could prove to be important in the development and qualification of any 2" generation reusable launch vehicle using composites as a structural element.

Engberg, Robert↗

Towards resolving protein structures at the atomic scale using atom probe tomography

In the field of Structural Biology, Atom Probe Tomography (APT) is in the nascent stages of development wherein coarse-grained visuals of proteins have been captured. The characterization of organic samples or biomolecules through the technique is currently limited to the detection of a few dominant signatures. The problem of indecipherable characterization can inherently be traced back to multiple forms of technique-specific responses to organic samples and consequent triggers leading to organic-sample and sample-medium interactions. While it is possible for captured manifestations of protein reconstructions to seemingly appear intact from a basic visual purview, the parameter-protein associative responses throughout the structure as a direct consequence of the inherent workings of the technique (until harmonized with organic sample complexity and behavior) and field evaporation-based factors make non-aberrative atomic associations infeasible. The work focused on identifying and theorizing the (above stated and other) fundamental mechanisms that stronghold the study of intricate atomic to higher order associations in proteins through APT. Attempts at structure elucidation of the cryogenic sample under study, through indirect associations (and methods) based on other imaging techniques, further revealed the distinct and highly distortive nature at the atomic scale deterring structural tunability and thus characterization of APT based cryogenic samples under analysis. As a direct counter to the atomic scale characterization problem, by taking the experiment-specific uncertainties, and probable APT-centric organic sample-based variabilities into account, a basic result is extracted and presented. Through mass-spectrometric and computational analysis, specific individual amino acids (Sulfur-containing protein-bound amino acids) in proteins and aspects of protein structure (probable backbone fragments, partial sequence - partial backbone portions) have been identified and characterized. Under analysis considerations, a few of the simplest known and easily inferable segments that favor structural deteriorations in the reconstructions are stated. Additionally, to overcome technique-specific deterrents to the characterization of biomolecules in cryogenic sample medium, the development of a protein-labeling strategy tailored to APT is suggested.

59 BASIC BIOLOGICAL SCIENCES↗

Structures of a synthetic antibody selected against and bound to the C‐terminal domain of Clostridium perfringens enterotoxin

Abstract Clostridium perfringensenterotoxin (CpE) causes cytotoxic gastrointestinal disease in mammalian epithelium by binding membrane protein receptors called claudins. Claudins direct the formation of cell/cell tight junctions through oligomerization and govern the transport of molecules between individual cells. CpE binds claudins through its C‐terminal domain (cCpE) and induces cytotoxicity through its N‐terminal domain. The non‐toxic cCpE is a useful tool to study claudins, tight junctions, and for translational applications, such as increasing the permeability of restrictive tissues like the blood–brain barrier or selective targeting of claudin overexpressing cancers. Conversely, there are no specialized molecular tools to study CpE or cCpE, or to modulate or inhibit their functions. We previously reported the development of synthetic antigen‐binding fragments (sFabs) that bind cCpE, and low‐resolution structures of them bound to claudin/cCpE complexes. Here, we determine high‐resolution structures of sFab COP‐2 bound to cCpE using X‐ray crystallography and cryogenic electron microscopy. The structures and biophysical findings provide the mechanism of COP‐2 binding to cCpE and the molecular determinants driving their interactions. These insights can advance the design of new antibody‐based tools from our COP‐2 scaffold to study or alter cCpE function and give rise to a “Trojan horse” strategy that exploits cCpE's tight junction barrier disrupting function to selectively deliver conjugated therapeutics through normally impermeable tissues.

Biochemistry & Molecular Biology↗

Durability Characterization of Advanced Polymeric Composites at Cryogenic Temperatures

The next generation of reusable launch vehicles will require technology development in several key areas. Of these key areas, the development of polymeric composite cryogenic fuel tanks promises to present one of the most difficult technical challenges. It is envisioned that a polymer matrix composite (PMC) tank would be a large shell structure capable of containing cryogenic fuels and carrying a range of structural loads. The criteria that will be imposed on such a design include reduced weight, conformal geometry, and impermeability. It is this last criterion, impermeability, that will provide the focus of this paper. The essence of the impermeability criterion is that the tank remains leak free throughout its design lifetime. To address this criterion, one of the first steps is to conduct a complete durability assessment of the PMC materials. At Langley Research Center, a durability assessment of promising new polyimide-based PMCs is underway. This durability program has focused on designing a set of critical laboratory experiments that will determine fundamental material properties under combined thermal-mechanical loading at cryogenic temperatures. The test program provides measurements of lamina and laminate properties, including strength, stiffness, and fracture toughness. The performance of the PMC materials is monitored as a function of exposure conditions and aging time. Residual properties after exposure are measured at cryogenic temperatures and provide quantitative values of residual strength and stiffness. Primary degradation mechanisms and the associated damage modes are measured with both destructive and nondestructive techniques. In addition to mechanical properties, a range of physical properties, such as weight, glass transition, and crack density, are measured and correlated with the test conditions. This paper will report on the progress of this research program and present critical results and illustrative examples of current findings.

Gates, T. S.↗

Fractional watt Vuillemier cryogenic refrigerator program engineering notebook. Volume 2: Stress analysis

A structural analysis performed on the 1/4-watt cryogenic refrigerator. The analysis covered the complete assembly except for the cooling jacket and mounting brackets. Maximum stresses, margin of safety, and natural frequencies were calculated for structurally loaded refrigerator components shown in assembly drawings. The stress analysis indicates that the design is satisfactory for the specified vibration environment, and the proof, burst, and normal operating loads.

Miller, W. S.↗