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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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10 records
All sourcesNASA NTRSDOE OSTI
NASA NTRS1976

Rotating Gravity Gradiometer Study

The application of a Rotating Gravity Gradiometer (RGG) system on board a Lunar Polar Orbiter (LPO) for the measurement of the Lunar gravity field was investigated. A data collection simulation study shows that a gradiometer will give significantly better gravity data than a doppler tracking system for the altitudes under consideration for the LOP, that the present demonstrated sensitivity of the RGG is adequate for measurement of the Lunar gravity gradient field, and that a single RGG instrument will provide almost as much data for geophysical interpretation as an orthogonal three axis RGG system. An engineering study of the RGG sensor/LPO spacecraft interface characteristics shows that the RGG systems under consideration are compatible with the present models of the LPO spacecraft.

Forward, R. L.↗
NASA NTRS1980

The development of an elastic reverse gradient garment to be used as a countermeasure for cardiovascular deconditioning

Using a new nomex lycra elastic fabric and individualized garment engineering techniques, reverse gradient garments (RGG's) were designed, constructed, and tested for effectiveness as a countermeasure against cardiovascular deconditioning. By combining torso compensated positive pressure breathing with a distally diminishing gradient of counterpressure supplied by the elastic fabric on the limbs, the RGG acts to pool blood in the extremities of recumbent persons much as though they were standing erect in 1 g. The RGG stresses the vasculature in a fashion similar to that experienced by the normally active man, hence preventing or limiting the development of post weightlessness orthostatic intolerance and related conditions. Four male, college age subjects received daily treatments with the RGG during a 15 day bedrest study. Four additional subjects also underwent the bedrest, but received no treatments; they served as controls. The preliminary indication was that the RGG was somewhat effective in limiting the deconditioning process.

Annis, J. F.↗
NASA NTRS1973

The development of an elastic reverse gradient garment to be used as a countermeasure for cardiovascular deconditioning

Using a new Nomex-Lycra elastic fabric and individualized garment engineering techniques, reverse gradient garments (RGG's) were designed, constructed, and tested for effectiveness as a countermeasure against cardiovascular deconditioning. By combining torso-compensated positive pressure breathing with a distally diminishing gradient of counterpressure supplied by the elastic fabric on the limbs, the RGG acts to pool blood in the extremities of recumbent persons much as though they were standing erect in 1 g. It was theorized that through the use of a dynamic pressurization scheme, the RGG would stress the vasculature in a fashion similar to that experienced by the noramlly active man, hence preventing or limiting the development of post-weightlessness orthostatic intolerance and related conditions. Four male, college-age subjects received daily treatments with the RGG during a 15-day bedrest study. Four additional subjects also underwent the bedrest, but received no treatments; they served as controls. The design and construction of the garments are described, and results of the treatment related measurements are given.

Annis, J. F.↗
NASA NTRS1982

Rotating gravity gradiometer study

Two rotating gravity gradiometer (RGG) sensors, along with all the external electronics needed to operate them, and the fixtures and special test equipment needed to fill and align the bearings, were assembled in a laboratory, and inspected. The thermal noise threshold of the RGG can be lowered by replacing a damping resistor in the first stage electronics by an active artificial resistor that generates less random voltage noise per unit bandwidth than the Johnson noise from the resistor it replaces. The artificial resistor circuit consists of an operational amplifier, three resistors, and a small DC to DC floating power supply. These are small enough to be retrofitted to the present circuit boards inside the RGG rotor in place of the 3 Megohm resistor. Using the artificial resistor, the thermal noise of the RGG-2 sensor can be lowered from 0.3 Eotvos to 0.15 Eotvos for a 10 sec integration time.

Forward, R. L.↗
DOE OSTI2020

SPLIT: Stable Protein Coacervation Using a Light Induced Transition

Protein coacervates serve as hubs to concentrate and sequester proteins and nucleotides and thus function as membraneless organelles to manipulate cell physiology. We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light. Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light. We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain. Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellularsized water-in-oil emulsions. An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae. Finally, the methods described here provide novel strategies for inducing protein phase separation using light.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2020

Identifying sequence perturbations to an intrinsically disordered protein that determine its phase-separation behavior

Phase separation of intrinsically disordered proteins (IDPs) commonly underlies the formation of membraneless organelles, which compartmentalize molecules intracellularly in the absence of a lipid membrane. Identifying the protein sequence features responsible for IDP phase separation is critical for understanding physiological roles and pathological consequences of biomolecular condensation, as well as for harnessing phase separation for applications in bioinspired materials design. To expand our knowledge of sequence determinants of IDP phase separation, we characterized variants of the intrinsically disordered RGG domain from LAF-1, a model protein involved in phase separation and a key component of P granules. Based on a predictive coarse-grained IDP model, we identified a region of the RGG domain that has high contact probability and is highly conserved between species; deletion of this region significantly disrupts phase separation in vitro and in vivo. We determined the effects of charge patterning on phase behavior through sequence shuffling. We designed sequences with significantly increased phase separation propensity by shuffling the wild-type sequence, which contains well-mixed charged residues, to increase charge segregation. This result indicates the natural sequence is under negative selection to moderate this mode of interaction. We measured the contributions of tyrosine and arginine residues to phase separation experimentally through mutagenesis studies and computationally through direct interrogation of different modes of interaction using all-atom simulations. Finally, we show that despite these sequence perturbations, the RGG-derived condensates remain liquid-like. Together, these studies advance our fundamental understanding of key biophysical principles and sequence features important to phase separation.

36 MATERIALS SCIENCE↗
DOE OSTI2021

Incorporation and Assembly of a Light-Emitting Enzymatic Reaction into Model Protein Condensates

Eukaryotic cells partition enzymes and other cellular components into distinct subcellular compartments to generate specialized biochemical niches. A subclass of these compartments form in the absence of lipid membranes, via liquid-liquid phase separation of proteins to form biomolecular condensates or “membraneless organelles” such as nucleoli, stress granules, and P-bodies. Because of their ability to form compartments from simple starting materials, membraneless organelles are an attractive target for engineering new functionalities in both living cells and protocells. In this work, we demonstrate incorporation of novel enzymatic activity in protein coacervates with a light-generating enzyme, NanoLuc, to produce bioluminescence. Using condensates comprised of the disordered RGG domain of C. elegans LAF-1, we show functionalization of condensates with enzymatic activity in vitro and that localization to protein coacervates enhances the assembly and activity of split enzymes. To build condensates that function as light emitting reactors, we designed a NanoLuc enzyme flanked by RGG domains. Furthermore, the resulting condensates concentrated NanoLuc by 10-fold over bulk solution and display significantly increased net reaction rates. We further show that condensate viscosity impacts light emission due to diffusion-limited behavior. By splitting NanoLuc enzyme into its constituent components, we demonstrate that NanoLuc activity can be reconstituted via co-condensation. Further, we demonstrate control of the spatial localization of enzyme within condensates by targettng NanoLuc to the surface of in vitro condensates. Collectively, this work demonstrates that membraneless organelles can be endowed with localized enzymatic activity, and that this activity can be spatially and temporally controlled via enzyme reconstitution and design of protein surfactants.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)

Mutations in the RNA-binding protein FUS cause familial amyotropic lateral sclerosis (ALS). Several mutations that affect the proline-tyrosine nuclear localization signal (PY-NLS) of FUS cause severe juvenile ALS. FUS also undergoes liquid–liquid phase separation (LLPS) to accumulate in stress granules when cells are stressed. In unstressed cells, wild type FUS resides predominantly in the nucleus as it is imported by the importin Karyopherin-β2 (Kapβ2), which binds with high affinity to the C-terminal PY-NLS of FUS. Here, we analyze the interactions between two ALS-related variants FUS(P525L) and FUS(R495X) with importins, especially Kapβ2, since they are still partially localized to the nucleus despite their defective/missing PY-NLSs. The crystal structure of the Kapβ2·FUS(P525L) PY-NLS complex shows the mutant peptide making fewer contacts at the mutation site, explaining decreased affinity for Kapβ2. Biochemical analysis revealed that the truncated FUS(R495X) protein, although missing the PY-NLS, can still bind Kapβ2 and suppresses LLPS. FUS(R495X) uses its C-terminal tandem arginine-glycine-glycine regions, RGG2 and RGG3, to bind the PY-NLS binding site of Kapβ2 for nuclear localization in cells when arginine methylation is inhibited. These findings suggest the importance of the C-terminal RGG regions in nuclear import and LLPS regulation of ALS variants of FUS that carry defective PY-NLSs.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1983

Evaluation of a Reverse Gradient Garment for prevention of bed-rest deconditioning

A Reverse Gradient Garment (RGG) was used to intermittently induce venous pooling in the extremities of a magnitude similar to that seen in going from a lying to standing position during the course of a 15-d period of horizontal bed rest. Venous pooling failed to improve bed-rest-induced losses in +2.5 Gz and +3.0 Gz centrifugation tolerance or to prevent increased heart-rate responses to lower-body negative pressure (LBNP). Four subjects served as controls, four were treated. Tests during the 7-d recovery period showed fluid/electrolyte and body composition values to have returned to pre-bed-rest levels with continued depression of acceleration tolerance times (56% decreased at +2.5 Gz and 74% decreased at +3.0 Gz compared to pre-bed-rest levels) and exaggerated blood insulin response on glucose tolerance testing (blood insulin for treated group increased 95% at 1 h before bed rest and 465% during recovery). This study demonstrates that the physiologic changes after bed rest persist for significant periods of time. Acceleration tolerance time proved to be a sensitive test for the deconditioning process.

Sandler, H.↗
DOE OSTI2021

Designer membraneless organelles sequester native factors for control of cell behavior

Subcellular compartmentalization of macromolecules increases flux and prevents inhibitory interactions to control biochemical reactions. Inspired by this functionality, we sought to build designer compartments that function as hubs to regulate the flow of information through cellular control systems. Here we report a synthetic membraneless organelle platform to control endogenous cellular activities through sequestration and insulation of native proteins. We engineer and express a disordered protein scaffold to assemble micron size condensates and recruit endogenous clients via genomic tagging with high-affinity dimerization motifs. By relocalizing up to ninety percent of a targeted enzymes to synthetic condensates, we efficiently control cellular behaviors, including proliferation, division, and cytoskeletal organization. Further, we demonstrate multiple strategies for controlled cargo release from condensates to switch cells between functional states. These synthetic organelles offer a powerful and generalizable approach to modularly control cell decision-making in a variety of model systems with broad applications for cellular engineering.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗