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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 721 records · Page 40
All sourcesNASA NTRSDOE OSTI
DOE OSTI2013

The usefulness of three-dimensional cell culture in induction of cancer stem cells from esophageal squamous cell carcinoma cell lines

Highlights: •Spheroids were created from esophageal carcinoma cells using NanoCulture® Plates. •The proportion of strongly ALDH-positive cells increased in 3-D culture. •Expression of cancer stem cell-related genes was enhanced in 3-D culture. •CA-9 expression was enhanced, suggesting hypoxia had been induced in 3-D culture. •Drug resistance was increased. 3-D culture is useful for inducing cancer stem cells. -- Abstract: In recent years, research on resistance to chemotherapy and radiotherapy in cancer treatment has come under the spotlight, and researchers have also begun investigating the relationship between resistance and cancer stem cells. Cancer stem cells are assumed to be present in esophageal cancer, but experimental methods for identification and culture of these cells have not yet been established. To solve this problem, we created spheroids using a NanoCulture® Plate (NCP) for 3-dimensional (3-D) cell culture, which was designed as a means for experimentally reproducing the 3-D structures found in the body. We investigated the potential for induction of cancer stem cells from esophageal cancer cells. Using flow cytometry we analyzed the expression of surface antigen markers CD44, CD133, CD338 (ABCG2), CD318 (CDCP1), and CD326 (EpCAM), which are known cancer stem cell markers. None of these surface antigen markers showed enhanced expression in 3-D cultured cells. We then analyzed aldehyde dehydrogenase (ALDH) enzymatic activity using the ALDEFLUOR reagent, which can identify immature cells such as stem cells and precursor cells. 3-D-cultured cells were strongly positive for ALDH enzyme activity. We also analyzed the expression of the stem cell-related genes Sox-2, Nanog, Oct3/4, and Lin28 using RT-PCR. Expression of Sox-2, Nanog, and Lin28 was enhanced. Analysis of expression of the hypoxic surface antigen marker carbonic anhydrase-9 (CA-9), which is an indicator of cancer stem cell induction and maintenance, revealed that CA-9 expression was enhanced, suggesting that hypoxia had been induced. Comparison of cancer drug resistance using cisplatin and doxorubicin in 3-D-cultured esophageal cancer cells showed that cancer drug resistance had increased. These results indicate that 3-D culture of esophageal squamous cell carcinoma lines is a useful method for inducing cancer stem cells.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2019

Three-dimensional explicit finite element formulation for shear localization with global tracking of embedded weak discontinuities

Shear localization is a damage mechanism characterized by the concentration of plastic deformation within thin bands of materials, often observed as a precursor to failure in metals. In this study, we present a three-dimensional explicit finite element formulation with embedded weak discontinuities for the treatment of shear localization under dynamic loading conditions. In this formulation, the onset of localization is detected via a material stability analysis suitable for rate-sensitive materials, and the continuity of propagating shear bands is ensured using a global tracking strategy involving a heat-conduction type boundary value problem, solved for a scalar level set function over the global domain of the problem. In addition, we propose a modified quadrature rule that is used to compute the contribution of an individual element to the global finite element arrays, taking into account the position of the embedded shear band within that element.

42 ENGINEERING↗
DOE OSTI2012

Effect of metal ions (Sn and Zn) on the thermal property of akaganeite nanorods

Graphical abstract: Compared with β-FeOOH and Zn doped β-FeOOH, the thermal stability of tin doped β-FeOOH increases both in solution and air. Zn and Sn ions doping favor the formation of rod like shape β-FeOOH. Highlights: ► The addition of tin ions increases the thermal stability of β-FeOOH nanorods both in solution and air. ► Tin ions doped β-FeOOH nanorods show higher decomposition temperature (ΔT = 200 °C). ► The presence of tin and zinc ions is favor of the formation of rod like shape. -- Abstract: The electrical, optical, and adsorption properties of akaganeite (β-FeOOH) can be modified by doping or combining with variety of elements. We prepared metal ions (Sn, Zn) doped β-FeOOH using β-FeOOH nanorods as the precursor by the hydrothermal method and investigated the effect of metal ions (Sn, Zn) on the thermal property of β-FeOOH nanorods. The doping of metal ions (Sn, Zn) was confirmed by X-ray powder diffraction, energy dispersive spectroscopy, and atomic emission spectrometer analysis. Thermogravimetric analysis and differential thermal analysis showed that they have different thermal decomposition temperatures. β-FeOOH nanorods and Zn doped β-FeOOH nanorods were transformed into α-Fe{sub 2}O{sub 3} particles at 400 °C. While Sn doped β-FeOOH nanorods should be heated to higher temperature (600 °C). Sn doped β-FeOOH nanorods showed higher thermal stability due to the Sn ions binding with the framework of β-FeOOH. Both Sn and Zn ions doped β-FeOOH remained the rod like shape.

36 MATERIALS SCIENCE↗
DOE OSTI2020

Formation and energetics of amorphous rare earth (RE) carbonates in the RE 2 O 3 –CO 2 –H 2 O system

Amorphous materials are of interest in the genesis of crystalline solids. They are often the first to precipitate from supersaturated solutions and are in the nanometer size range, transforming into a nanocrystalline metastable phase, and then to thermodynamically stable phases. One must seek knowledge of the composition, structure, thermodynamics, kinetics, and reaction pathways that relate the amorphous and crystalline phases. The subject of this study is the amorphous phases in the system RE 2 O 3 -CO 2 –H 2 O (RE- La, Nd, Dy,Yb). They were synthesized via direct precipitation and urea hydrolysis, and characterized by powder XRD, DTA with mass spectrometric analysis of the evolved gases. Phases have the non-stoichiometric composition RE 2 O 3 ·xCO 2 ·yH 2 O (1 < x < 3, 3 < y <7), which is different from that of crystalline simple carbonates and hydroxycarbonates. Thus we consider them to be amorphous precursors, rather than amorphous carbonates. High temperature oxide melt solution calorimetry in molten sodium molybdate solvent was used to derive their enthalpies of formation from oxides and elements. Increase of energetic stability per mole of RE 2 O 3 compounds occurs in the order: RE 2 O 3 → 2RE(OH)3→ 2REO(OH) → RE 2 O 2 CO 3 → Amorphous precursor → 2(REOHCO 3 ) → RE 2 (CO 3 ) 3 ·yH 2 O. We calculated the enthalpies of possible transformations of amorphous precursors to crystalline phases and conclude that thermodynamics determines the dependence of the crystallizing products on the temperature and partial pressure of CO 2 and H 2 O. Amorphous precursors are clearly intermediate in the synthesis, and ternary phases containing both H 2 O and CO 2 compete with each other in terms of thermodynamic stability. The energy landscape obtained here will allow one to directly synthesize specific products and control their functionality.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2016

Mechanism of conductivity relaxation in liquid and polymeric electrolytes: Direct link between conductivity and diffusivity

Combining broadband impedance spectroscopy, differential scanning calorimetry, and nuclear magnetic resonance we analyzed charge and mass transport in two polymerized ionic liquids and one of their monomeric precursors. In order to establish a general procedure for extracting single-particle diffusivity from their conductivity spectra, we critically assessed several approaches previously employed to describe the onset of diffusive charge dynamics and of the electrode polarization in ion conducting materials. Based on the analysis of the permittivity spectra, we demonstrate that the conductivity relaxation process provides information on ion diffusion and the magnitude of cross-correlation effects between ionic motions. A new approach is introduced which is able to estimate ionic diffusivities from the characteristic times of conductivity relaxation and ion concentration without any adjustable parameters. Furthermore, this opens the venue for a deeper understanding of charge transport in concentrated and diluted electrolyte solutions.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗
DOE OSTI2021

Influence of biogenic emissions from boreal forests on aerosol–cloud interactions

Boreal forest acts as a carbon sink and contributes to the formation of secondary organic aerosols via emission of aerosol precursor compounds. However, these influences on the climate system are poorly quantified. Here we show direct observational evidence that aerosol emissions from the boreal forest biosphere influence warm cloud microphysics and cloud-aerosol interactions in a scale-dependent and highly dynamic manner. Analyses of in situ and ground-based remote-sensing observations from the SMEAR II station in Finland, conducted over eight months in 2014, reveal substantial increases in aerosol load over the forest one to three days after aerosol-poor marine air enters the forest environment. We find that these changes are consistent with secondary organic aerosol formation and, together with water-vapour emissions from evapotranspiration, are associated with changes in the radiative properties of warm, low-level clouds. Here, the feedbacks between boreal forest emissions and aerosol-cloud interactions and the highly dynamic nature of these interactions in air transported over the forest over timescales of several days suggest boreal forests have the potential to mitigate climate change on a continental scale. Our findings suggest that even small changes in aerosol precursor emissions, whether due to changing climatic or anthropogenic factors, may substantially modify the radiative properties of clouds in moderately polluted environments. Emissions from the boreal forest biosphere can substantially increase aerosol load above the forest and influence the radiative properties of clouds, according to analysis of observations from a monitoring station in Finland.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2016

Evolutionary analysis of iron (Fe) acquisition system in Marchantia polymorpha

Summary To acquire appropriate iron (Fe), vascular plants have developed two unique strategies, the reduction‐based strategy I of nongraminaceous plants for Fe 2+ and the chelation‐based strategy II of graminaceous plants for Fe 3+ . However, the mechanism of Fe uptake in bryophytes, the earliest diverging branch of land plants and dominant in gametophyte generation is less clear. Fe isotope fractionation analysis demonstrated that the liverwort Marchantia polymorpha uses reduction‐based Fe acquisition. Enhanced activities of ferric chelate reductase and proton ATPase were detected under Fe‐deficient conditions. However, M. polymorpha did not show mugineic acid family phytosiderophores, the key components of strategy II, or the precursor nicotianamine. Five ZIP (ZRT/IRT‐like protein) homologs were identified and speculated to be involved in Fe uptake in M. polymorpha . MpZIP3 knockdown conferred reduced growth under Fe‐deficient conditions, and MpZIP3 overexpression increased Fe content under excess Fe. Thus, a nonvascular liverwort, M. polymorpha , uses strategy I for Fe acquisition. This system may have been acquired in the common ancestor of land plants and coopted from the gametophyte to sporophyte generation in the evolution of land plants.

Lo, Jing‐Chi↗
DOE OSTI2018

Enhancing metabolic flux to photosynthetic biofuels

The overall objective of this project was to test new metabolic engineering and systems biology approaches for maximizing metabolic flux to biofuels in cyanobacterial hosts. Our central hypothesis was that combining directed metabolic engineering approaches with global rewiring of circadian transcriptional programs would lead to enhanced biofuel productivities. As proof of principle, we applied our novel toolset to optimize cyanobacterial production of isobutyraldehyde (IBA), which is a direct precursor of isobutanol, starting from a parental IBA-producing strain of Synechococcus elongatus PCC 7942.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Development of an Integrated Mechanistic Source Term Assessment Capability for Lead- and Sodium-Cooled Fast Reactors

The scope of this project entails development of mechanistic source term assessment capabilities to inform and guide plant design activities and to support license applications. The main objective of this project is to couple NL’s SAS4A safety analysis code with the FAI’s FATE facility modeling code for mechanistic source term assessments in liquid-metal-cooled fast reactors. The end product is a coupled SAS4A-FATE code system for integrated mechanistic source term assessments and Level-2 PRA as the precursor for off-site dispersion analyses (Level-3 PRA). The SAS4A code is used to model system transients with fuel failures and its results are linked to the FATE code to predict radionuclide transport through the reactor vessel cover gas and containment system. The integrated code package offers a state-of-the-art radionuclide tracking capability for a spectrum of accident scenarios consistent with the plant dynamic response including the reactivity feedback to support Probabilistic Risk Assessments (PRA).

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗
DOE OSTI2015

Fine precipitation scenarios of AlZnMg(Cu) alloys revealed by advanced atomic-resolution electron microscopy study Part II: Fine precipitation scenarios in AlZnMg(Cu) alloys

Although they are among the most important precipitation-hardened materials for industry applications, the high-strength AlZnMg(Cu) alloys have thus far not yet been understood adequately about their underlying precipitation scenarios in relation with the properties. This is partly due to the fact that the structures of a number of different precipitates involved in the alloys are unknown, and partly due to the complexity that the precipitation behaviors of the alloys may be closely related to the alloy's composition. In Part I of the present study, we have determined all the unknown precipitate structures in the alloys. Here in Part II, using atomic-resolution electron microscopy in association with the first principles energy calculations, we further studied and correlated the phase/structure transformation/evolution among these hardening precipitates in relation with the alloy's composition. It is shown that there are actually two coexisting classes of hardening precipitates in these alloys: the first class includes the η′-precipitates and their early-stage Guinier–Preston (GP-η′) zones; the second class includes the precursors of the equilibrium η-phase (referred to η{sub p}, or η-precursor) and their early-stage Guinier–Preston (GP-η{sub p}) zones. The two coexisting classes of precipitates correspond to two precipitation scenarios. - Highlights: • We determine and verify all the key precipitate structures in AlMgZn(Cu) alloys. • We employ aberration-corrected scanning transmission electron microscopy (STEM). • We use aberration-corrected high-resolution TEM (HRTEM) for the investigations. • We obtain atomic-resolution images of the precipitates and model their structures. • We refine all precipitate structures with quantitative image simulation analysis. • The hardening precipitates in AlZnMg alloys shall be classified into two groups. • Two precipitation scenarios coexist in the alloys. • The precipitation behavior of such an alloy depends on the alloy's composition. • Very detailed phase/structure transformations among the precipitates are revealed.

36 MATERIALS SCIENCE↗
DOE OSTI2015

Rapid Analysis of Microalgal Triacylglycerols with Direct-Infusion Mass Spectrometry

Cultivation of microalgae has the potential to provide lipid-derived feedstocks for conversion to liquid transportation fuels. Lipid extracts from microalgae are significantly more complex than those of traditional seed oils, and their composition changes significantly throughout the microalgal growth period. With three acyl side chains per molecule, triglycerides (TAGs) are an important fuel precursor, and the distribution of acyl chain composition for TAGs has a significant impact on fuel properties and processing. Therefore, determination of the distribution of microalgal TAG production is needed to assess the value of algal extracts designed for fuel production and to optimize strain, cultivation, and harvesting practices. Methods utilized for TAG speciation commonly involve complicated and time-consuming chromatographic techniques. Here we present a method for TAG speciation and quantification based on direct-infusion mass spectrometry, which provides rapid characterization of TAG profiles without chromatographic separation. Specifically, we utilize Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to provide a reference library of TAGs for the microalgae Nannochloropsis sp. that provides the basis for high-throughput TAG quantitation by time-of-flight mass spectrometry (TOF MS). In conclusion, we demonstrate the application of this novel approach for lipid characterization with respect to TAG compound distribution, which informs both immediate and future strain and process optimization strategies.

09 BIOMASS FUELS↗
DOE OSTI2014

A metabonomic evaluation of the monocrotaline-induced sinusoidal obstruction syndrome (SOS) in rats

The main curative treatment of colorectal cancer remains the surgery. However, when metastases are suspected, surgery is followed by a preventive chemotherapy using oxaliplatin which, unfortunately, may cause liver sinusoidal obstruction syndrome (SOS). Such hepatic damage is barely detected during or after chemotherapy due to a lack of effective diagnostic procedures, but liver biopsy. The primary objective of the present study was to identify potential early diagnosis biomarkers of SOS using a metabonomic approach. SOS was induced in rats by monocrotaline, a prototypical toxic substance. {sup 1}H NMR spectroscopy analysis of urine samples collected from rats treated with monocrotaline showed significant metabolic changes as compared to controls. During a first phase, cellular protective mechanisms such as an increased synthesis of GSH (reduced taurine) and the recruitment of cell osmolytes in the liver (betaine) were seen. In the second phase, the disturbance of the urea cycle (increased ornithine and urea reduction) leading to the depletion of NO, the alteration in the GSH synthesis (increased creatine and GSH precursors (glutamate, dimethylglycine and sarcosine)), and the liver necrosis (decrease taurine and increase creatine) all indicate the development of SOS. - Highlights: • Urine metabonomic profiles of SOS have been identified. • Urine osmoprotectants and anti-oxidants indicated an initial liver protection. • Liver necrosis was demonstrated by increased urine levels of taurine and creatine. • NO depletion was suggested by changes in ornithine and urea.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2025

Delamination-informed lifecycle decisions: A dielectric and machine learning framework for composite sorting and recycling

Composite materials are widely used in aerospace, marine, and automotive sectors due to their high strength-to-weight ratio and durability. However, their long-term reliability can be compromised by damage accumulation. Specifically, delamination initiation serves as a precursor to structural failure, which is often difficult to detect during damage inspection. Identifying and sorting delamination initiation in samples not only increases operational safety while providing critical information for end-of-life decisions, which influences both the service life extension value and the efficiency of fiber extraction during recycling. This research addresses two challenges: (1) developing a nondestructive, ex-situ framework to sort composite materials based on damage severity, particularly delamination, and (2) understanding how damage in composites influences resin removal during pyrolysis. Both experimental work and finite element analysis were performed to predict critical stress levels that are associated with delamination onset. Based on these results, three loading levels 50 %, 75 %, and 90 % of maximum stress, were selected for controlled experiments, generating composite samples with varying extents of damage for machine learning model training. Microscopic imaging of these samples confirmed the damage progression from matrix cracking to delamination, validating the computational predictions. We explored supervised machine learning using dielectric measurements to classify damage states. Preliminary results show an artificial neural network can identify early delamination which is a potential precursor to failure, with 94.44 % accuracy on our dataset. A parallel investigation into the effect of damage severity on pyrolysis recycling showed that heavily delaminated samples required significantly less energy for comparable matrix removal than undamaged samples.

dielectric variables↗
DOE OSTI2021

Mixed Tin-Titanium Oxides by Atomic Layer Deposition on Planar Substrates: Physical and Electronic Structure

Ternary atomic layer deposition of Sn-doped TiO x (Sn:TiO x ) and Ti-doped SnO x (Ti:SnO x ) was performed using tetrakisdimethylamidotitanium (IV) and tin(IV) with water at 150 °C on (1 0 0) Si and quartz substrates. The physical structure of the films was investigated using scanning electron, atomic force, and Kelvin probe force microscopies as well as spectroscopic ellipsometry, Raman spectroscopy, and X-ray diffraction. All as-deposited films are amorphous and possessed flat and conformal topographies with low surface roughness (<1 nm). Annealed Sn:TiO x and Ti:SnO x crystallize into the tetragonal anatase TiO 2 and cassiterite SnO 2 structures, respectively, without large variations to the film topography. The chemical composition of the as-deposited and annealed films was investigated through X-ray photoelectron spectroscopy. While dopants are homogenously mixed, the ternary ALD to form Sn:TiO x and Ti:SnO x surprisingly does not follow the rule of mixtures, despite similarities in both the growth rates of binary SnO x and TiO x and the precursors used. This finding is also supported by spectroscopic ellipsometry. Here, we postulate the deviation from the rule of mixtures is due to non-ideal surface chemistry of the binary ALD of SnO x . The optical bandgap of all films was quantified by the Tauc analysis and the valence band position was investigated by valence band X-ray photoelectron spectroscopy. Intra-bandgap states above the valence band maximum were observed for as-deposited Ti:SnO x and Sn:TiO x and ascribed to 5 s states from Sn 2+ ions. The presence of these intra-bandgap states induces enhanced visible absorption and bathochromic shifts to the apparent bandgap for all as-deposited films. Annealing the films removes the intra-bandgap states and increases the optical bandgap. Simplified band diagrams created using the data available demonstrated that the conduction and valence band positions did not vary between the positions of pure TiO 2 or SnO 2 with monotonic increases to the dopant incorporation.

36 MATERIALS SCIENCE↗
DOE OSTI2022

Vertical gradient freeze growth of detector grade CdZnTeSe single crystals

Here, we report the growth of detector grade Cd 0.9 Zn 0.1 Te 0.97 Se 0.03 (CZTS) single crystals, a recently discovered quaternary semiconductor for room temperature radiation detection, by a vertical gradient freeze (VGF) method. VGF is a comparatively low-temperature growth method and avoids relative motion between the heater and the ampoule containing the precursor materials which minimizes any thermal drift or temperature fluctuations. As a result, CZTS single crystals with superior charge transport properties has been obtained. Growth of detector-grade CZTS single crystals using VGF method has not been reported yet. X-ray spectroscopy based elemental analysis showed that the grown crystals demonstrated the desired stoichiometry required for high resolution radiation detection. Planar detectors fabricated by deposition of gold contacts (~0.07 cm 2 ) demonstrated high bulk resistivity ~10 10 Ω-cm and a very low leakage current density of 2.8 × 10 –8 A/cm 2 at a bias of 100 V when measured at room temperature. The detectors showed excellent radiation response with 100 % charge collection efficiency when exposed to 5486 keV alpha particles. The electron mobility-lifetime (μτ) product was measured to be 3 × 10 –3 cm 2 /V using a single polarity Hecht analysis which is at par with the recently reported values measured in CZTS grown using conventional Bridgman or travelling heater method. The electron mobility has been calculated to be 964 cm 2 V –1 s –1 using a time-of-flight (TOF) method, a substantial improvement over that obtained from conventionally grown CZTS single crystals.

36 MATERIALS SCIENCE↗
DOE OSTI2014

Preparation and application of zirconium sulfate supported on SAPO-34 molecular sieve as solid acid catalyst for esterification

Graphical abstract: - Highlights: • SAPO-34 supported zirconium sulfate solid acid catalyst was prepared. • Esterification of acetic acid with ethanol can be catalyzed by ZS/SAPO-34. • The hydration of ZS is vital to the acidic property and catalytic performance. • The ZS/SAPO-34 catalyst treated at 200 °C shows good reusability. - Abstract: Zirconium sulfate (ZS) was supported on SAPO-34 molecular sieve by using an incipient wetness impregnation method with zirconium sulfate as the precursor. The as-prepared catalysts were used as solid acid catalyst for esterification reaction of acetic acid with ethanol. The influence of calcination temperature on the acidic property, catalytic activity, and reusability of ZS/SAPO-34 catalysts were mainly investigated. FT-IR, SEM, EDS and TG analysis have been carried out to demonstrate the characteristics of ZS/SAPO-34 catalysts. It was found that the 30 wt%ZS/SAPO-34 catalysts display the property of superacid irrespective of calcination temperature. The ZS/SAPO-34 catalyst treated at 200 °C can enhance the interaction between the supported ZS and SAPO-34 and keep the catalyst remaining substantially active after several reaction cycles. However, further increasing calcination temperature will cause the transfer of ZS from hydrate to anhydrous phase, and thus the decrease of activity.

36 MATERIALS SCIENCE↗
DOE OSTI2023

Multiphysics Analysis of the MSRE Experiment Using Griffin-SAM Coupled Code System

Molten salt reactors (MSRs) with flowing fuel have the unique feature of utilizing the fuel salt for heat generation and extraction at the same time since the fuel salt is circulating through the whole primary loop of the reactor. This movement of the fuel salt results in a partial decay of the delayed neutron precursors (DNPs) outside the core and corresponding redistribution in the active core region. To capture this phenomenon accurately, neutronics and thermal hydraulics computational tools need to be modified to handle the movement of the DNPs and their decay. Idaho and Argonne National Laboratories are actively working on developing neutronics and thermal hydraulics tools to model MSRs with flowing fuel. The neutronics code Griffin and the thermal hydraulics code SAM were extended to handle flowing fuel with the drift of the DNPs under the MOOSE framework. In this work, a simplified Multiphysics model of the Molten Salt Reactor Experiment (MSRE) is developed and utilized to perform steady-state and transient analyses. The MSRE experiments were designed to show the impact of the DNPs losses on core reactivity. The following section provides a description of the MSRE experiment with the core main parameters used to develop the model. Then, the developed Multiphysics model of the MSRE is presented. Finally, steady-state verification tests and unprotected loss of flow transient tests are discussed followed by a summary and conclusions.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2018

Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes

Membrane protein characterization is consistently hampered by challenges with expression, purification, and solubilization. Among several biophysical techniques employed for their characterization, native-mass spectrometry (MS) has emerged as a powerful tool for the analysis of membrane proteins and complexes. Here, two MS platforms, the FT-ICR and Q-ToF, have been explored to analyze the homotetrameric water channel protein, AquaporinZ (AqpZ), under non-denaturing conditions. This 97 kDa membrane protein complex can be readily liberated from the octylglucoside (OG) detergent micelle under a range of instrument conditions on both MS platforms. Increasing the applied collision energy of the FT-ICR collision cell yielded varying degrees of tetramer (97 kDa) liberation from the OG micelles, as well as dissociation into the trimeric (72 kDa) and monomeric (24 kDa) substituents. Tandem-MS on the Q-ToF yielded higher intensity tetramer signal and, depending on the m/z region selected, the observed monomer signal varied in intensity. Precursor ion selection of an m/z range above the expected protein signal distribution, followed by mild collisional activation, is able to efficiently liberate AqpZ with a high S/N ratio. The tetrameric charge state distribution obtained on both instruments demonstrated superpositioning of multiple proteoforms due to varying degrees of N-terminal formylation. .

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗