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At least 217 records · Page 12

Facets control charge separation during photoelectrochemical water oxidation with strontium titanate (SrTiO 3 ) single crystals

The photocatalytic overall water splitting reaction provides a pathway to hydrogen fuel from sunlight. Photocatalysts must achieve the reaction without the application of an external bias, which requires an effective charge separation mechanism. Photolabeling studies and electrostatic simulations for the well-known CoOOH/Al:SrTiO 3 /Rh/Cr 2 O 3 photocatalyst suggest that charge separation is driven by work function differences at the (100) and (110) facets of SrTiO 3 , which are electron and hole selective, respectively. Here we use hydrogen annealed SrTiO 3–x single crystals to obtain the first quantitative assessment of the charge separation ability of the (100), or (110), or (111) facets during oxygen evolution. Under UV illumination (60 mW cm –2 ), the crystals exhibit variable water oxidation photocurrents (0.34, 0.82, 1.36 mA cm –2 at 1.23 V versus RHE) and photovoltage values of 1.40, 1.52 and 1.52 V for (100), (110), and (111) SrTiO 3–x , respectively. A surface photovoltage increase in that same order (0.31 V < 0.57 V < 0.67 V) is confirmed independently with vibrating Kelvin probe surface photovoltage spectroscopy (VKP-SPV) under 375 nm (1.91 mW cm –2 ) illumination. Mott Schottky measurements in aqueous K 3/4 [Fe(CN) 6 ] reveal facet-dependent flatband positions of –0.58, –0.71, and –0.74 V RHE for the (100), (110), and (111) crystals respectively. This confirms that the photoelectrochemical water oxidation performance of SrTiO 3–x crystals is controlled by the work function of each facet, which determines the electron transfer barrier height of the respective solid–liquid junctions. After correcting for differences in electron donor concentrations, barriers are found to increase in the order (100) < (111) < (110) and differ by as much as 0.16 eV, similar to an earlier prediction. Altogehter, these results explain the charge separation mechanism in SrTiO 3 photocatalysts and highlight the need for faceted semiconductor crystals as light absorbers in particle-based photocatalysts.

08 HYDROGEN↗

Technology Case Study: Economic, Sustainability, and Deployment Considerations for Sustainable Aviation Fuels Produced via Lignocellulosic Sugar Catalysis

This report presents a technology case study reflecting one exemplary representative pathway for the conversion of lignocellulosic sugars to sustainable aviation fuels (SAF) via aqueous phase reforming (APR) catalysis, considered within a broader integrated biorefinery framework based on biochemical processing operations. While far from the only option for converting sugars to SAF, this pathway was selected as a case study here based on its relatively high technology maturity and simplistic processing approach (avoiding complex separations or other equipment scalability challenges), coupled with the potential for high fuel yields and favorable costs/carbon intensities with opportunities for further near-term optimization. The report considers key process integration and engineering design considerations for a modeled hypothetical, nth-plant commercial biorefinery, reflecting a number of processing options and parameters envisioned to be achievable as future goals. Resultant outputs from Aspen Plus process simulations are evaluated through techno-economic and life cycle analyses (TEA and LCA), including implications for marginal cost of CO2 abatement and inclusion of currently-applicable policy incentives. Moving beyond base case configurations, a number of alternative scenarios are also evaluated for their ability to further improve economics, greenhouse gas (GHG) emissions, and marginal cost of abatement, highlighting a path to achieve deep decarbonization goals of more than 70% GHG reduction for SAF (with the potential to reach net-negative carbon intensities in some cases) under reasonable fuel production costs. The report also highlights future opportunities and gaps for further research on this technology pathway.

09 BIOMASS FUELS↗

Valorizing Hydrothermal liquefaction aqueous phase via nanofiltration: Enhancing biocrude production from algal biomass

The hydrothermal liquefaction aqueous phase (HTL-AP), a nutrient- and carbon-rich waste byproduct of the HTL process, can be recirculated to enhance biocrude production efficiency. To avoid dilution of the HTL feedstock and reduced fuel quality associated with recirculating untreated HTL-AP, this study investigated nanofiltration to concentrate organic compounds and separate nutrients from HTL-AP of three algal biomass sources: an algal wastewater treatment system and two from harmful algal blooms in lakes. Up to 99% of the organics were recovered in the concentrated HTL-AP (retentate), and significant increases in both permeate flux and permeation of nitrogenous compounds were achieved by increasing HTL-AP temperature to 45 °C and pH to 11. Recirculating the concentrated HTL-AP increased biocrude yield by 70%-116% and reduced biocrude nitrogen content by up to 12%. The carbon and energy capture in the biocrude also increased by 66% and 68%, respectively.

09 BIOMASS FUELS↗

Photoinduced Carbon Dioxide Release via a Metastable Photoacid in a Nonaqueous Environment

Capturing and concentrating carbon dioxide (CO 2 ) from the atmosphere is an important modern scientific and technological challenge. CO 2 can be captured by forming a carbamate bond with amines, most notably monoethanolamine (MEA). Regenerating MEA by releasing the captured CO 2 requires heating the carbamate solution, which is energy-demanding and wasteful. Recently, photoacids were used to convert light input into pH change, which in turn released CO 2 from aqueous carbonate solutions. Here, we report a merocyanine photoacid that releases CO 2 from a non-aqueous carbamate solution of MEA, thereby extending the utility of this concept to a wider range of capture agents and environments. Based on the absorption spectra of the photoacid in the presence of acids and CO 2 , we show that the photoacid cycle and the CO 2 capture of MEA are two separate equilibria that are coupled to each other via protons. Then, we demonstrate that irradiating 405 nm light on the sample can induce release of CO 2 which we detect using an in-line mass spectrometer. This work highlights an alternative path for optimizing a photo-induced CO 2 capture and release system.

Amines↗

Method of cross-linking polyvinyl alcohol and other water soluble resins

A self supporting sheet structure comprising a water soluble, noncrosslinked polymer such as polyvinyl alcohol which is capable of being crosslinked by reaction with hydrogen atom radicals and hydroxyl molecule radicals is contacted with an aqueous solution having a pH of less than 8 and containing a dissolved salt in an amount sufficient to prevent substantial dissolution of the noncrosslinked polymer in the aqueous solution. The aqueous solution is then irradiated with ionizing radiation to form hydrogen atom radicals and hydroxyl molecule radicals and the irradiation is continued for a time sufficient to effect crosslinking of the water soluble polymer to produce a water insoluble polymer sheet structure. The method has particular application in the production of battery separators and electrode envelopes for alkaline batteries.

Phillipp, W. H.↗

An electrochemical rebalance cell for Redox systems

An electrochemical rebalance cell for maintaining electrochemical balance, at the system level, of the acidified aqueous iron chloride and chromium chloride reactant solutions in the redox energy storage system was constructed and evaluated. The electrochemical reaction for the cathode is Fe(+3) + e(-) yields Fe(+2), and that for the anode is 1/2H2 yields H(+) + e(-). The iron (carbon felt) electrode and the hydrogen (platinized carbon) electrode are separated by an anion exchange membrane. The performance of the rebalance cell is discussed as well as the assembly of a single rebalance cell and multicell stacks. Various cell configurations were tested and the results are presented and discussed. The rebalance cell was also used to demonstrate its ability, as a preparative tool, for making high purity solutions of soluble reduced metal ionic species. Preparations of titanium, copper, vanadium and chromium ions in acidified solutions were evaluated.

Acevedo, J. C.↗

Climate studies with a multilayer energy balance model. III - Climatic impact of stratospheric volcanic aerosols

A multilayer energy balance model is applied in an examination of the sensitivity of climate to stratospheric aerosols induced by volcanic eruptions. Zonally and annually averaged quantities are considered, with ocean and land temperatures computed separately and the atmosphere below the 200 mb level divided into eight layers of 24 sublayers each. The aerosol is assumed to form in the 150-200 mb range. Aerosol parameters for radiative transfer calculations are reflection in the solar spectral region and absorption in the solar and IR regions. A 75 percent aqueous solution of sulfuric acid is assumed for the aerosols. The sensitivity of the hemispherically averaged surface temperature is enhanced 37 percent, with a 20 percent uncertainty, when the thermal IR radiation is excluded. The solar radiation enhances the surface temperatures to a higher degree than the thermal radiation. The maximum response to the evenly distributed aerosols is in the 60-70 deg N latitudes and propagates, weakening, to lower latitudes.

Chou, M.-D.↗

On the Relationship between Cosmic Ray Exposure Ages and Petrography of CM Chondrites

Carbonaceous (C) chondrites are potentially the most primitive among chondrites because they mostly escaped thermal metamorphism that affected the other chondrite groups. C chondrites are chemically distinguished from other chondrites by their high Mg/Si ratios and refractory elements, and have experienced various degrees of aqueous alteration. They are subdivided into eight subgroups (CI, CM, CO, CV, CK, CR, CB and CH) based on major element and oxygen isotopic ratios. Their elemental ratios vary over a wide range, in contrast to those of ordinary and enstatite chondrites which are relatively uniform. It is critical to know how many separate bodies are represented by the C chondrites. In this study we defined 4 distinct cosmic-ray exposure (CRE) age groups of CMs and systematically characterized the petrography in each of the 4 CRE age groups to determine whether the groups have significant petrographic differences with such differences probably reflecting different parent body (asteroid) geological processing, or multiple original bodies. We have reported the results of a preliminary grouping at the NIPR Symp. in 2013 [3], however, we revised the grouping and here report our new results.

Takenouchi, A.↗

Key Recent Scientific Results from the Opportunity Rover's Exploration of Endeavour Crater, Mars

The Opportunity Rover is currently in its 11th year of operations, exploring the rim of the approximately 22 km wide Noachian-age Endeavour Crater. Opportunity spent its 5th winter season in Cook Haven, a gentle swale along Murray Ridge. Two small rocks serendipitously overturned by rover wheel motions show evidence for aqueous precipitation of sulfates, and interaction with a strong oxidant (e.g., O2) to form a thin, high valence state Mn oxide coating. After the winter, Opportunity headed south to Cape Tribulation and explored Shoemaker formation impact breccias, finding numerous Ca-sulfate veins cutting across outcrops. A key target for Opportunity's measurements has been the Spirit of Saint Louis crater (SoSL), which is approximately 25 m wide, oval in plan view, shallow, flat-floored, and has a slightly raised rim. SoSL crater is surrounded by an apron of bright, polygonally-shaped outcrops and is superimposed on a gentle swale in Cape Tribulation. Rocks in a thin reddish zone on the rim are enriched in hematite, Si, and Ge, and depleted in Fe, relative to surrounding rocks. Apron rocks include an outcrop also enriched in Si and Ge, and slightly depleted in Fe. In general rocks in the crater and apron have elevated S relative to Shoemaker formation breccias, tracking values observed in the Cook Haven and the Hueytown (fracture running perpendicular to Cape Tribulation) outcrops. SoSL crater lies just to the west of Marathon Valley, a key target for exploration by Opportunity because five separate CRISM observations indicate the presence of Fe/Mg smectites on the upper valley floor. Opportunity data show that low relief, relatively bright polygonal outcrops dominate the valley floor where not covered by scree and soil shed from surrounding walls. Initial reconnaissance shows that the outcrops are breccias with compositions similar to the typical SoSL crater apron and floor rocks, although only the very upper portion of the valley has been explored as of August 2015. Pervasive but modest aqueous alteration of Endeavour's rim is implied by the combination of CRISM and Opportunity data, providing insight into early aqueous processes dominated in this location by relatively low water to rock ratios, and at least in part associated with enhanced fluid flow along fractures.

Arvidson, R. E.↗

Structure–Function Relationships in Sequence-Controlled Copolymers for Rare Earth Element Chelation

The ability to tune material function through primary sequence is a defining feature of biological macromolecules, allowing precise control over structure and target interactions in complex aqueous environments. However, translating sequence–structure–function relationships to synthetic macromolecules is challenging due to their dispersity in sequence, conformation, and composition. Here, we report systematic studies of amphiphilic polymer chelators designed to probe how composition and patterning influence binding affinity and selectivity for rare earth elements (REEs), a series of technologically relevant metals with challenging separation profiles. A library of copolymers varying hydrophobic monomer composition and patterning was synthesized via reversible addition–fragmentation chain transfer (RAFT) polymerization, spanning statistical, gradient, and block architectures. REE binding was quantified using a high-throughput colorimetric assay, and reconstruction of polymer ensembles using kinetic stochastic simulations enabled quantitative comparisons of sequence heterogeneity, linking local monomer colocalization to emergent REE binding. Further, we investigated the role of different hydrophobic comonomers in tuning metal coordination, with binding trends linked to structural features that influence binding site desolvation. Complementary dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS) measurements showed that both polymer and monomer architecture modulate metal-induced conformational changes, and that multichain assembly behavior emerges beyond critical hydrophobic thresholds. Sequence control also altered REE selectivity, with nonmonotonic differences observed across compositionally identical polymers with different sequence architectures. Together, these findings establish design principles that connect polymer sequence and structure to binding performance, guiding the design of macromolecular chelators with enhanced affinity and selectivity for applications in separations, sensing, and catalysis.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Lunar and Planetary Science XXXV: Mars: Remote Sensing and Terrestrial Analogs

The session "Mars: Remote Sensing and Terrestrial Analogs" included the following:Physical Meaning of the Hapke Parameter for Macroscopic Roughness: Experimental Determination for Planetary Regolith Surface Analogs and Numerical Approach; Near-Infrared Spectra of Martian Pyroxene Separates: First Results from Mars Spectroscopy Consortium; Anomalous Spectra of High-Ca Pyroxenes: Correlation Between Ir and M ssbauer Patterns; THEMIS-IR Emissivity Spectrum of a Large Dark Streak near Olympus Mons; Geomorphologic/Thermophysical Mapping of the Athabasca Region, Mars, Using THEMIS Infrared Imaging; Mars Thermal Inertia from THEMIS Data; Multispectral Analysis Methods for Mapping Aqueous Mineral Depostis in Proposed Paleolake Basins on Mars Using THEMIS Data; Joint Analysis of Mars Odyssey THEMIS Visible and Infrared Images: A Magic Airbrush for Qualitative and Quantitative Morphology; Analysis of Mars Thermal Emission Spectrometer Data Using Large Mineral Reference Libraries ; Negative Abundance : A Problem in Compositional Modeling of Hyperspectral Images; Mars-LAB: First Remote Sensing Data of Mineralogy Exposed at Small Mars-Analog Craters, Nevada Test Site; A Tool for the 2003 Rover Mini-TES: Downwelling Radiance Compensation Using Integrated Line-Sight Sky Measurements; Learning About Mars Geology Using Thermal Infrared Spectral Imaging: Orbiter and Rover Perspectives; Classifying Terrestrial Volcanic Alteration Processes and Defining Alteration Processes they Represent on Mars; Cemented Volcanic Soils, Martian Spectra and Implications for the Martian Climate; Palagonitic Mars: A Basalt Centric View of Surface Composition and Aqueous Alteration; Combining a Non Linear Unmixing Model and the Tetracorder Algorithm: Application to the ISM Dataset; Spectral Reflectance Properties of Some Basaltic Weathering Products; Morphometric LIDAR Analysis of Amboy Crater, California: Application to MOLA Analysis of Analog Features on Mars; Airborne Radar Study of Soil Moisture at a Mars Analog Site: Tohachi Wash/Little Colorado River; and Antarctic Dry Valleys: Modification of Rocks and Soils and Implications for Mars The Arkaroola Mars Analogue Region, South Australia.

Source record↗

An Evaluation of the Vapor Phase Catalytic Ammonia Removal Process for Use in a Mars Transit Vehicle

An experimental program has been developed to evaluate the potential of the Vapor Phase Catalytic Ammonia Reduction (VPCAR) technology for use as a Mars Transit Vehicle water purification system. Design modifications which will be required to ensure proper operation of the VPCAR system in reduced gravity are also evaluated. The VPCAR system is an integrated wastewater treatment technology that combines a distillation process with high temperature catalytic oxidation. The distillation portion of the system utilizes a vapor compression distillation process to provide an energy efficient phase change separation. This portion of the system removes any inorganic salts and large molecular weight, organic contaminates, i.e., non-volatile, from the product water stream and concentrates these contaminates into a byproduct stream. To oxidize the volatile organic compounds and ammonia, a vapor phase, high temperature catalytic oxidizer is used. This catalytic system converts these compounds along with the aqueous product into CO2, H2O, and N2O. A secondary catalytic bed can then be used to reduce the N2O to nitrogen and oxygen (although not evaluated in this study). This paper describes the design specification of the VPCAR process, the relative benefits of its utilization in a Mars Transit Vehicle, and the design modification which will be required to ensure its proper operation in reduced gravity. In addition, the results of an experimental evaluation of the processors is presented. This evaluation presents the processors performance based upon product water purity, water recovery rates, and power.

Flynn, Michael↗

Surfactant-like peptide gels are based on cross-β amyloid fibrils

Surfactant-like peptides, in which hydrophilic and hydrophobic residues are encoded within different domains in the peptide sequence, undergo facile self-assembly in aqueous solution to form supramolecular hydrogels. These peptides have been explored extensively as substrates for the creation of functional materials since a wide variety of amphipathic sequences can be prepared from commonly available amino acid precursors. The self-assembly behavior of surfactant-like peptides has been compared to that observed for small molecule amphiphiles in which nanoscale phase separation of the hydrophobic domains drives the self-assembly of supramolecular structures. Here, we investigate the relationship between sequence and supramolecular structure for a pair of bola-amphiphilic peptides, Ac-KLIIIK-NH 2 (L2) and Ac-KIIILK-NH 2 (L5). Despite similar length, composition, and polar sequence pattern, L2 and L5 form morphologically distinct assemblies, nanosheets and nanotubes, respectively. Cryo-EM helical reconstruction was employed to determine the structure of the L5 nanotube at near-atomic resolution. Rather than displaying self-assembly behavior analogous to conventional amphiphiles, the packing arrangement of peptides in the L5 nanotube displayed steric zipper interfaces that resembled those observed in the structures of β-amyloid fibrils. Like amyloids, the supramolecular structures of the L2 and L5 assemblies were sensitive to conservative amino acid substitutions within an otherwise identical amphipathic sequence pattern. This study highlights the need to better understand the relationship between sequence and supramolecular structure to facilitate the development of functional peptide-based materials for biomaterials applications.

Das, Abhinaba [Emory University, Atlanta, GA (Unit↗

Developing Digital Twin Visualizations: A Methodology and Case Study on Chemical Separation Processing

As advances in digital engineering continue to push the technological boundaries, digital twin (DT) visualizations for diagnostics and safeguards advancement become much more feasible and practical. DTs generate large and complex data streams that require effective user interfaces to provide monitoring and diagnostic capabilities. Unfortunately, while these frameworks exist, there is not much research on the systematic documentation of human–computer interaction (HCI) for DT visualization. This work presents a dual-mode visualization methodology (two dimensional [2D] graphical user interface dashboard and 3D mixed reality) designed to support diagnostic tasks in DT systems and building on a validated framework and applying established HCI principles. The methodology is demonstrated through a case study of aqueous processing at Idaho National Laboratory, using experimental data from the chemical solvent extraction runs. Our interfaces display real-time alerts and monitoring to inform users of safeguards anomalies. The interfaces use immersive 3D mixed-reality visualization for further system and experiment investigation. This work demonstrates how the systematic application of HCI principles can inform DT visualization design for diagnostic and safeguards applications. While formal user evaluation studies remain as future work, this paper documents the systematic design methodology and demonstrates a proof-of-concept implementation.

3D visualization↗

Formation of thin sulfide rinds on the CM parent body

Many of the textures that are observed in CM chondrites have been alternately assigned nebular, parent-body, or combined nebular-parent body origins. Since it is very difficult to substantiate the production of complex textures in the nebula, an alternate approach to this problem is to rigorously determine which of the observable textures could have been produced by reasonable aqueous alteration processes on the CM parent body. Potential parent body reactions involving S deserve special attention because S-bearing phase define many important CM textures, such as rims. We have examined the possibility that the thin (about 5 microns) rinds of sulfides observed around some partially dissolved olivines within the chondrules and matrixes of CM chondrites were formed by the preferential precipitation of sulfides at or near dissolving olivine boundaries during parent-body alteration. Our model defines two infinite and parallel planes of olivine that are separated by pure water. Average separation distances between olivine grains in CM chondrites at the time of accretion (about 100-200 microns) were estimated by assuming a closed system fluid/rock ratio of 45% and varying the bimodal grain-size distribution. We restrict our calculations to the case of an isochemical system with sufficient bisulfide in solution to account for precipitation of pyrrhotite at STP. Our model examines the possibility that dissolving olivines with compositions between Fo(sub 0.9) and Fo(sub 0.4) can produce a strong gradient of Fe(2+) at pHs from 7 to 12 such that the precipitation of pyrrhotite will be initiated at the olivine-fluid boundary. Since CM phase equilibria is consistent with highly reducing conditions, Fe released by olivines would largely remain in solution as Fe(2+) until the solubility product of a Fe(2+) bearing phase was exceeded. Our calculations indicate that all examined combinations of olivine composition and pH yield a strong Fe(2+) gradient at the time the pyrrhotite solubility is exceeded near the olivine-fluid boundary. This favors the initiation of sulfide precipitation around dissolving olivines. However, sulfide rinds are not observed around all partially dissolved olivine grains in CM chondrites, which suggests that the availability of aqueous sulfide species limits the production of the thin sulfide rinds.

Browning, L. B.↗

Materials processing in space

Processing-refining of raw materials from extraterrestrial sources is detailed for a space materials handling facility. The discussion is constrained to those steps necessary to separate desired components from raw or altered input ores, semi-purified feedstocks, or process scrap and convert the material into elements, alloys, and consumables. The materials are regarded as originating from dead satellites and boosters, lunar materials, and asteroids. Strong attention will be given to recycling reagent substances to avoid the necessity of transporting replacements. It is assumed that since no aqueous processes exist on the moon, the distribution of minerals will be homogeneous. The processing-refining scenario will include hydrochemical, pyrochemical, electrochemical, and physical techniques selected for the output mass rate/unit plant mass ratio. Flow charts of the various materials processing operations which could be performed with lunar materials are provided, noting the necessity of delivering several alloying elements from the earth due to scarcities on the moon.

Waldron, R. D.↗

Lunar and Planetary Science XXXVI, Part 6

Contents include the following: A Model for Multiple Populations of Presolar Diamonds. Characterization of Martian North Polar Geologic Units Using Mars Odyssey THEMIS Data. Effect of Flow on the Internal Structure of the Martian North Polar Layered Deposits. Elemental Abundance Distributions in Basalt Clays and Meteorites: Is It a Biosignature? Early Results on the Saturn System from the Composite Infrared Spectrometer. NanoSIMS D/H Imaging of Isotopically Primitive Interplanetary Dust Particles. Presolar (Circumstellar and Interstellar) Phases in Renazzo: The Effects of Parent Body Processing. Catastrophic Disruption of Hydrated Targets: Implications for the Hydrated Asteroids and for the Production of Interplanetary Dust Particles. Chemical and Mineralogical Analyses of Particles from the Stratospheric Collections Coinciding with the 2002 Leonid Storm and the 2003 Comet Grigg-Skjellerup Trail Passage. An Analysis of the Solvus in the CaS-MnS System. ESA s SMART-1 Mission at the Moon: First Results, Status and Next Steps. Europa Analog Ice-splitting Measurements and Experiments with Ice-Hunveyor on the Frozen Balaton-Lake, Hungary. Chromium on Eros: Further Evidence of Ordinary Chondrite Composition. Dust Devil Tracks on Mars: Observation and Analysis from Orbit and the Surface. Spatial Variation of Methane and Other Trace Gases Detected on Mars: Interpretation with a General Circulation Model. Mars Water Ice and Carbon Dioxide Seasonal Polar Caps: GCM Modeling and Comparison with Mars Express Omega Observations. Component Separation of OMEGA Spectra with ICA. Clathrate Formation in the Near-Surface Environment of Titan. Space Weathering: A Proposed Laboratory Approach to Explaining the Sulfur Depletion on Eros. Sample Collection from Small Airless Bodies: Examination of Temperature Constraints for the TGIP. Sample Collector for the Hera Near-Earth Asteroid Sample Return Mission. A Rugged Miniature Mass-Spectrometer for Measuring Aqueous Geochemistry on Mars. Martian and Lunar Pyroxene Microstructures Studied by Single-Crystal X-Ray Diffraction.

Source record↗

Distribution and Spectroscopy of Green Fluorescent Protein and Acyl-CoA: Cholesterol Acytransferase in Sf21 Insect Cells

Acyl-CoA: cholesterol acyltransferase (ACAT) is thought to significantly participate in the pathway of cholesterol esterification that underlies the pathology of artherosclerosis. This enzyme is a membrane protein known to be preferentially bound within the endoplasmic reticulum of mammalian cells, from which location it esterifies cholesterol derived from low density lipoprotein. Cultures of insect cells were separately infected with baculovirus containing the gene for green fluroescent protein (GFP) and with baculovirus containing tandem genes for GFP and ACAT. These infected cultures expressed GFP and the fusion protein GCAT, respectively, with maximum expression occurring on the fourth day after infection. Extraction of GFP- and of GCAT-expressing cells with urea and detergent resulted in recovery of fluorescent protein in aqueous solution. Fluorescence spectra at neutral pH were identical for both GFP and GCAT extracts in aqueous solution, indicating unperturbed tertiary structure for the GFP moiety within GCAT. In a cholesterol esterification assay, GCAT demonstrated ACAT activity, but with less efficiency compared to native ACAT. It was hypothesized that the membrane protein ACAT would lead to differences in localization of GCAT compared to GFP within the respective expressing insect cells. The GFP marker directly and also within the fusion protein GCAT was accordingly used as the intracellular probe that was fluorescently analyzed by the new biophotonics technique of hyperspectral imaging. In that technique, fluorescence imaging was obtained from two dimensional arrays of cells, and regions of interest from within those images were then retrospectively analyzed for the emission spectra that comprises the image. Results of hyperspectral imaging of insect cells on day 4 postinfection showed that GCAT was preferentially localized to the cytoplasm of these cells compared to GFP. Furthermore, the emission spectra obtained for the localized GCAT displayed a peak blue shift from 518nm obtained in neutral aqueous solution to 505nm obtained in localized regions within the cells. This blue shift indicates change in the fluorescence coupling of the GFP moiety of GCAT. It is hypothesized that change in tertiary environment of GCAT, coincident with intracellular deposition of GCAT, follows from intracellular trafficking of GCAT leading to membrane interactions with the ACAT moiety, and/or self-assembly of GCAT, that alters the chromophore environment of the GFP moiety of GCAT. These findings introduce a new technique of biophotonic imaging to studies of intracellular protein trafficking and interactions. This technique of hyperspectral imaging could contribute to advancing the emergent field of proteomics. Because of the noninvasive nature of this technique, kinetic processes associated with intracellular protein trafficking, and interactions of proteins within cellular domains, can be considered for investigation within a single cell as well as a cell population.

Richmond, R. C.↗