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At least 577 records · Page 32

Kainate receptor channel opening and gating mechanism

Kainate receptors, a subclass of ionotropic glutamate receptors, are tetrameric ligand-gated ion channels that mediate excitatory neurotransmission. Kainate receptors modulate neuronal circuits and synaptic plasticity during the development and function of the central nervous system and are implicated in various neurological and psychiatric diseases, including epilepsy, depression, schizophrenia, anxiety and autism. Although structures of kainate receptor domains and subunit assemblies are available, the mechanism of kainate receptor gating remains poorly understood. Here we present cryo-electron microscopy structures of the kainate receptor GluK2 in the presence of the agonist glutamate and the positive allosteric modulators lectin concanavalin A and BPAM344. Concanavalin A and BPAM344 inhibit kainate receptor desensitization and prolong activation by acting as a spacer between the amino-terminal and ligand-binding domains and a stabilizer of the ligand-binding domain dimer interface, respectively. Channel opening involves the kinking of all four pore-forming M3 helices. Our structures reveal the molecular basis of kainate receptor gating, which could guide the development of drugs for treatment of neurological disorders.

59 BASIC BIOLOGICAL SCIENCES↗

Spatial proteomics reveals signal sequence characteristics correlated with localization in cyanobacteria

Abstract Cyanobacteria have an inner and outer cell membrane enclosing the periplasm and cell wall and an additional set of internal membranes (called the thylakoid membranes) enclosing the thylakoid lumen. The periplasm and thylakoid lumen have unique proteomes, but the mechanisms regulating protein sorting to these locations have remained elusive. Here, proximity-based proteomics using the engineered peroxidase APEX2 was performed in the cyanobacteria Synechococcus sp. PCC 7002 to profile the proteomes of the cytoplasm, thylakoid lumen, and the periplasm and outer membrane (P-OM). Our analyses revealed specific roles for the thylakoid lumen in photosynthesis and energy generation, as well as roles for the periplasm in metabolite transport and binding, cell motility, and cell wall maintenance. Forty proteins localized to both the thylakoid lumen and the P-OM; however, their biological functions remain unclear. We also analyzed the correlation between signal sequence characteristics and differential protein localization to either the thylakoid lumen or the P-OM. In PCC 7002, as well as Synechocystis sp. PCC 6803 and Nostoc sp. PCC 7120, thylakoid lumen proteins translocated across membranes via the Secretory (Sec) system possessed more hydrophobic and alpha-helical signal sequence H-regions than P-OM proteins. The signal sequences of homologous proteins in Gloeobacter violaceus PCC 7421, a cyanobacterial species with a combined thylakoid lumen and periplasmic space, did not exhibit such differences. Therefore, the pattern of increased H-region hydrophobicity and alpha helix content is specific to cyanobacteria with a separate thylakoid lumen space and likely contributes to proper protein sorting between the thylakoid lumen and periplasm.

Plant Sciences↗

Angstrom-Resolved Interfacial Structure in Buried Organic-Inorganic Junctions

Charge transport processes at interfaces which are governed by complex interfacial electronic structure play a crucial role in catalytic reactions, energy storage, photovoltaics, and many biological processes. Here, the first soft X-ray second harmonic generation (SXR-SHG) interfacial spectrum of a buried interface (boron/Parylene-N) is reported. SXR-SHG shows distinct spectral features that are not observed in X-ray absorption spectra, demonstrating its extraordinary interfacial sensitivity. Comparison to electronic structure calculations indicates a boron-organic separation distance of 1.9 {\AA}, wherein changes as small as 0.1 {\AA} result in easily detectable SXR-SHG spectral shifts (ca. 100s of meV). As SXR-SHG is inherently ultrafast and sensitive to individual atomic layers, it creates the possibility to study a variety of interfacial processes, e.g. catalysis, with ultrafast time resolution and bond specificity.

74 ATOMIC AND MOLECULAR PHYSICS↗

Comparative transcriptomics uncovers poplar and fungal genetic determinants of ectomycorrhizal compatibility

Ectomycorrhizal symbiosis supports tree growth and is crucial for nutrient cycling and temperate and boreal ecosystems functioning. The establishment of functional ectomycorrhiza (ECM) first requires the association of compatible partners. However, host and fungal genetic determinants governing mycorrhizal compatibility are unknown. To identify such factors in poplar and its fungal associates, we mined existing and de novo tree and fungal transcriptional datasets. We identified co-expressed genes enabling ECM symbiosis at early and mature stages of the interaction. These sets of genes can be divided into general fungal-sensing and ECM-specific components. We highlight the importance of fungal modulation of plant JA-related defenses and the regulation of secretory pathways for ECM compatibility, including upregulation of key fungal small secreted proteins, the downregulation of plant secreted peroxidases, and the downregulation of plant cell wall remodeling proteins concomitantly with the upregulation of fungal glycosyl hydrolases acting on pectin. Not only gene regulation, but also its temporal scale and dynamics seem to play a crucial role for mycorrhizal compatibility. The expression profile of the host Common Symbiosis Pathway and nutrient transporters was also studied, revealing constitutive levels of expression and moderate upregulation in compatible ECM interactions. Overall, these results underscore the importance of novel biological functions during the establishment of ECM symbiosis, help us gain insights into the molecular events determining mycorrhiza compatibility, and serve as a data-rich transcriptomic resource to open new research questions in the field.

Marqués‐Gálvez, José Eduardo↗

Preliminary analyses of space radiation protection for lunar base surface systems

Radiation shielding analyses are performed for candidate lunar base habitation modules. The study primarily addresses potential hazards due to contributions from the galactic cosmic rays. The NASA Langley Research Center's high energy nucleon and heavy ion transport codes are used to compute propagation of radiation through conventional and regolith shield materials. Computed values of linear energy transfer are converted to biological dose-equivalent using quality factors established by the International Commision of Radiological Protection. Special fluxes of heavy charged particles and corresponding dosimetric quantities are computed for a series of thicknesses in various shield media and are used as an input data base for algorithms pertaining to specific shielded geometries. Dosimetric results are presented as isodose contour maps of shielded configuration interiors. The dose predictions indicate that shielding requirements are substantial, and an abbreviated uncertainty analysis shows that better definition of the space radiation environment as well as improvement in nuclear interaction cross-section data can greatly increase the accuracy of shield requirement predictions.

Nealy, John E.↗

Characterization of auxin-binding proteins from zucchini plasma membrane

We have previously identified two auxin-binding polypeptides in plasma membrane (PM) preparations from zucchini (Cucurbita pepo L.) (Hicks et al. 1989, Proc. Natl. Acad. Sci. USA 86, 4948-4952). These polypeptides have molecular weights of 40 kDa and 42 kDa and label specifically with the photoaffinity auxin analog 5-N3-7-3H-IAA (azido-IAA). Azido-IAA permits both the covalent and radioactive tagging of auxin-binding proteins and has allowed us to characterize further the 40-kDa and 42-kDa polypeptides, including the nature of their attachment to the PM, their relationship to each other, and their potential function. The azido-IAA-labeled polypeptides remain in the pelleted membrane fraction following high-salt and detergent washes, which indicates a tight and possibly integral association with the PM. Two-dimensional electrophoresis of partially purified azido-IAA-labeled protein demonstrates that, in addition to the major isoforms of the 40-kDa and 42-kDa polypeptides, which possess isoelectric points (pIs) of 8.2 and 7.2, respectively, several less abundant isoforms that display unique pIs are apparent at both molecular masses. Tryptic and chymotryptic digestion of the auxin-binding proteins indicates that the 40-kDa and 42-kDa polypeptides are closely related or are modifications of the same polypeptide. Phase extraction with the nonionic detergent Triton X-114 results in partitioning of the azido-IAA-labeled polypeptides into the aqueous (hydrophilic) phase. This apparently paradoxical behavior is also exhibited by certain integral membrane proteins that aggregate to form channels. The results of gel filtration indicate that the auxin-binding proteins do indeed aggregate strongly and that the polypeptides associate to form a dimer or multimeric complex in vivo. These characteristics are consistent with the hypothesis that the 40-kDa and 42-kDa polypeptides are subunits of a multimeric integral membrane protein which has an auxin-binding site, and which may possess transporter or channel function.

NASA Program Space Biology↗

Evidence for a role in growth and salt resistance of a plasma membrane H+-ATPase in the root endodermis

The plasma membrane of plant cells is energized by an electrochemical gradient produced by P-type H+-ATPases (proton pumps). These pumps are encoded by at least 12 genes in Arabidopsis. Here we provide evidence that isoform AHA4 contributes to solute transport through the root endodermis. AHA4 is expressed most strongly in the root endodermis and flowers, as suggested by promoter-GUS reporter assays. A disruption of this pump (aha4-1) was identified as a T-DNA insertion in the middle of the gene (after VFP(574)). Truncated aha4-1 transcripts accumulate to approximately 50% of the level observed for AHA4 mRNA in wild-type plants. Plants homozygous for aha4-1 (-/-) show a subtle reduction in root and shoot growth compared with wild-type plants when grown under normal conditions. However, a mutant phenotype is very clear in plants grown under salt stress (e.g., 75 or 110 mM NaCl). In leaves of mutant plants subjected to Na stress, the ratio of Na to K increased 4-5-fold. Interestingly, the aha4-1 mutation appears to be semidominant and was only partially complemented by the introduction of additional wild-type copies of AHA4. These results are consistent with the hypothesis that aha4-1 may produce a dominant negative protein or RNA that partially disrupts the activity of other pumps or functions in the root endodermal tissue, thereby compromising the function of this cell layer in controlling ion homeostasis and nutrient transport.

NASA Discipline Plant Biology↗

Hydrogen consumption by methanogens on the early Earth

It is possible that the first autotroph used chemical energy rather than light. This could have been the main source of primary production after the initial inventory of abiotic organic material had been depleted. The electron acceptor most readily available for use by this first chemoautotroph would have been CO2. The most abundant electron donor may have been H2 that would have been outgassing from volcanoes at a rate estimated to be as large as 10(12) moles yr-1, as well as from photo-oxidation of Fe+2. We report here that certain methanogens will consume H2 down to partial pressures as low as 4 Pa (4 x 10(-5) atm) with CO2 as the sole carbon source at a rate of 0.7 ng H2 min-1 microgram-1 cell protein. The lower limit of pH2 for growth of methanogens can be understood on the basis that the pH2 needs to be high enough for one ATP to be synthesized per CO2 reduced. The pH2 values needed for growth measured here are consistent with those measured by Stevens and McKinley for growth of methanogens in deep basalt aquifers. H2-consuming autotrophs are likely to have had a profound effect on the chemistry of the early atmosphere and to have been a dominant sink for H2 on the early Earth after life began rather than escape from the Earth's atmosphere to space.

Non-NASA Center↗

Production regimes in four eastern boundary current systems

High productivity (maxima ~3 g C m(sup -2)day(sup -1)) of the Eastern Boundary Currents (EBCs), i.e. the California, Peru-Humboldt, Canary and Benguela Currents, is driven by a combination of local forcing and large-scale circulation. The characteristics of the deep water brought to the surface by upwelling favorable winds depend on the large-scale circulation patterns. Here we use a new hydrographic and nutrient climatology together with satellite measurements ofthe wind vector, sea-surface temperature (SST), chlorophyll concentration, and primary production modeled from ocean color to quantify the meridional and seasonal patterns of upwelling dynamics and biological response. The unprecedented combination of data sets allows us to describe objectively the variability for small regions within each current and to characterize the governing factors for biological production. The temporal and spatial environmental variability was due in most regions to large-scale circulation, alone or in combination with offshore transport (local forcing). The observed meridional and seasonal patterns of biomass and primary production were most highlycorrelated to components representing large-scale circulation. The biomass sustained by a given nutrient concentration in the Atlantic EBCs was twice as large as that of the Pacific EBCs. This apparent greater efficiency may be due toavailability of iron, physical retention, or differences in planktonic community structure.

circulation patternsoceans↗

Regional to Global Assessments of Phytoplankton Dynamics From The SeaWiFS Mission

Photosynthetic production of organic matter by microscopic oceanic phytoplankton fuels ocean ecosystems and contributes roughly half of the Earth's net primary production. For 13 years, the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) mission provided the first consistent, synoptic observations of global ocean ecosystems. Changes in the surface chlorophyll concentration, the primary biological property retrieved from SeaWiFS, have traditionally been used as a metric for phytoplankton abundance and its distribution largely reflects patterns in vertical nutrient transport. On regional to global scales, chlorophyll concentrations covary with sea surface temperature (SST) because SST changes reflect light and nutrient conditions. However, the oceanmay be too complex to be well characterized using a single index such as the chlorophyll concentration. A semi-analytical bio-optical algorithm is used to help interpret regional to global SeaWiFS chlorophyll observations from using three independent, well-validated ocean color data products; the chlorophyll a concentration, absorption by CDM and particulate backscattering. First, we show that observed long-term, global-scale trends in standard chlorophyll retrievals are likely compromised by coincident changes in CDM. Second, we partition the chlorophyll signal into a component due to phytoplankton biomass changes and a component caused by physiological adjustments in intracellular chlorophyll concentrations to changes in mixed layer light levels. We show that biomass changes dominate chlorophyll signals for the high latitude seas and where persistent vertical upwelling is known to occur, while physiological processes dominate chlorophyll variability over much of the tropical and subtropical oceans. The SeaWiFS data set demonstrates complexity in the interpretation of changes in regional to global phytoplankton distributions and illustrates limitations for the assessment of phytoplankton dynamics using chlorophyll retrievals alone.

phytoplankton↗

Growth as an Alternative Approach to the Construction of Extra-Terrestrial Habitats

A critical component of human space exploration and eventual settlement is the ability to construct habitats while minimizing payload mass launched from Earth. To respond to this challenge, we have proposed the use of fungal bio-composites for ‘growing’ extra-terrestrial structures, directly at the destination, significantly lowering the mass of structural materials transported from Earth and minimizing the need for high mass robotic operations and infrastructure preparations. Throughout human history, the construction of habits has used biologically produced materials, from bone and skins to wood and limestone. Traditionally, the materials are used only after they die. Currently, the idea of working with living biological organisms, and the phenomenon of growth itself, is increasing in interest both, in architecture and space applications. Here we describe the use of mycelium-based composites as an alternative, biological approach for constructing regenerative and adaptive extraterrestrial habitats, a continuation of our research program initiated under the auspices of the ‘Myco-architecture Off Planet’ NASA NIAC Team. These composites, which are fire-resistant, insulating, do not consist of volatile organic compounds from petrochemical products and can be used independently or in conjunction with regolith, could employ the living biological growth in a controlled environment, for the process of material fabrication, assembly, maintenance, and repair, providing structures resilient to extra-terrestrial hazards. The paper will outline the potential and challenges of using bio-composites for space applications and will present how these might be addressed, in order to make this biological approach feasible, providing new, growing materials for design habitats on long-duration missions.

Monika Brandić Lipińska↗

Real-time bioelectronic sensing of environmental contaminants

Real-time chemical sensing is crucial for applications in environmental and health monitoring. Biosensors can detect a variety of molecules through genetic circuits that use these chemicals to trigger the synthesis of a coloured protein, thereby producing an optical signal. However, the process of protein expression limits the speed of this sensing to approximately half an hour, and optical signals are often difficult to detect in situ. Here we combine synthetic biology and materials engineering to develop biosensors that produce electrical readouts and have detection times of minutes. Here we programmed Escherichia coli to produce an electrical current in response to specific chemicals using a modular, eight-component, synthetic electron transport chain. As designed, this strain produced current following exposure to thiosulfate, an anion that causes microbial blooms, within 2 min. This amperometric sensor was then modified to detect an endocrine disruptor. The incorporation of a protein switch into the synthetic pathway and encapsulation of the bacteria with conductive nanomaterials enabled the detection of the endocrine disruptor in urban waterway samples within 3 min. Our results provide design rules to sense various chemicals with mass-transport-limited detection times and a new platform for miniature, low-power bioelectronic sensors that safeguard ecological and human health.

59 BASIC BIOLOGICAL SCIENCES↗

Increased expression of fatty acid and ABC transporters enhances seed oil production in camelina

Abstract Background Lipid transporters play an essential role in lipid delivery and distribution, but their influence on seed oil production in oilseed crops is not well studied. Results Here, we examined the effect of two lipid transporters, FAX1 ( fatty acid export1 ) and ABCA9 ( ATP-binding cassette transporter subfamily A9 ) on oil production and lipid metabolism in the oilseed plant Camelina sativa . Overexpression (OE) of FAX1 and ABCA9 increased seed weight and size, with FAX1 -OEs and ABCA9 -OEs increasing seed length and width, respectively, whereas FAX1 / ABCA9 -OEs increasing both. FAX1- OE and ABCA9- OE displayed additive effects on seed oil content and seed yield. Also, OE of FAX1 and ABCA9 affected membrane lipid composition in developing pods, especially on phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol. The expression of some genes involved in seed oil synthesis, such as DGAT2 , PDAT1 , and LEC1 , was increased in developing seeds of FAX1 - and/or ABCA9 -OEs. Conclusion These results indicate that increased expression of FAX1 and ABCA9 can potentially be applied to improving camelina oil production.

59 BASIC BIOLOGICAL SCIENCES↗

Phase partitioning, crystal growth, electrodeposition and cosmic ray experiments in microgravity

Five experiments are contained in one Get Away Special Canister (5 cu ft). The first utilizes microgravity to separate biological cells and to study the mechanism of phase partitioning in 12 separate cuvettes. Two experiments are designed to grow organic crystals by physical vapor transport. One experiment consists of eight electroplating cells with various chemicals to produce surfaces electroplated in microgravity. Some of the surfaces have micron sized particles of hard materials co-deposited during electrodeposition. The fifth experiment intercepts cosmic ray particles and records their paths on photographic emulsions. The first four experiments are controlled by an on-board C-MOS controller. The fifth experiment is totally passive. These are the first in Space. Their purpose is to create new commercial products with microgravity processing.

Wessling, Francis C.↗

NASA Strategy to Safely Live and Work in the Space Radiation Environment

In space, astronauts are constantly bombarded with energetic particles. The goal of the National Aeronautics and Space Agency and the NASA Space Radiation Project is to ensure that astronauts can safely live and work in the space radiation environment. The space radiation environment poses both acute and chronic risks to crew health and safety, but unlike some other aspects of space travel, space radiation exposure has clinically relevant implications for the lifetime of the crew. Among the identified radiation risks are cancer, acute and late CNS damage, chronic and degenerative tissue decease, and acute radiation syndrome. The term "safely" means that risks are sufficiently understood such that acceptable limits on mission, post-mission and multi-mission consequences can be defined. The NASA Space Radiation Project strategy has several elements. The first element is to use a peer-reviewed research program to increase our mechanistic knowledge and genetic capabilities to develop tools for individual risk projection, thereby reducing our dependency on epidemiological data and population-based risk assessment. The second element is to use the NASA Space Radiation Laboratory to provide a ground-based facility to study the health effects/mechanisms of damage from space radiation exposure and the development and validation of biological models of risk, as well as methods for extrapolation to human risk. The third element is a risk modeling effort that integrates the results from research efforts into models of human risk to reduce uncertainties in predicting the identified radiation risks. To understand the biological basis for risk, we must also understand the physical aspects of the crew environment. Thus, the fourth element develops computer algorithms to predict radiation transport properties, evaluate integrated shielding technologies and provide design optimization recommendations for the design of human space systems. Understanding the risks and determining methods to mitigate the risks are keys to a successful radiation protection strategy.

Cucinotta, Francis A.↗

A Strategy to Safely Live and Work in the Space Radiation Environment

The goal of the National Aeronautics and Space Agency and the Space Radiation Project is to ensure that astronauts can safely live and work in the space radiation environment. The space radiation environment poses both acute and chronic risks to crew health and safety, but unlike some other aspects of space travel, space radiation exposure has clinically relevant implications for the lifetime of the crew. The term safely means that risks are sufficiently understood such that acceptable limits on mission, post-mission and multi-mission consequences (for example, excess lifetime fatal cancer risk) can be defined. The Space Radiation Project strategy has several elements. The first element is to use a peer-reviewed research program to increase our mechanistic knowledge and genetic capabilities to develop tools for individual risk projection, thereby reducing our dependency on epidemiological data and population-based risk assessment. The second element is to use the NASA Space Radiation Laboratory to provide a ground-based facility to study the understanding of health effects/mechanisms of damage from space radiation exposure and the development and validation of biological models of risk, as well as methods for extrapolation to human risk. The third element is a risk modeling effort that integrates the results from research efforts into models of human risk to reduce uncertainties in predicting risk of carcinogenesis, central nervous system damage, degenerative tissue disease, and acute radiation effects. To understand the biological basis for risk, we must also understand the physical aspects of the crew environment. Thus the fourth element develops computer codes to predict radiation transport properties, evaluate integrated shielding technologies and provide design optimization recommendations for the design of human space systems. Understanding the risks and determining methods to mitigate the risks are keys to a successful radiation protection strategy.

Corbin, Barbara J.↗

Preliminary Analysis of a Nuclear Grade Sandwich Composite for Transportable Micro-Reactor Shielding

Transportable microreactors pose a unique shielding challenge compared with conventional reactors. Many vendors are seeking to transport a microreactor to a site, operate it for a number of years, and then transport it to a secondary site. This involves ensuring the reactor has sufficient shielding during operations and post-irradiation transportation to ensure the safety of those around it, all while ensuring it can be transported via traditional means. This work examines the use of a nuclear grade sandwich composite (NGSC) which seeks to combine the reactor pressure vessel and biological shielding into a single unit. Through this work, it was determined that the NGSC was sufficient to adequately reduce neutron and gamma current exiting the structure to warrant further investigation as a potential shielding option for microreactors.

42 ENGINEERING↗

Biocatalyst discovery and design for plastics deconstruction: A multi‐scale perspective

Plastic waste accumulation poses significant environmental challenges due to a lack of economical solutions for the molecular deconstruction of diverse synthetic polymers. Biological‐based degradation offers promise but is hindered by the crystallinity, hydrophobicity, and additive complexity of plastics, which restrict biocatalyst access and activity. To address these problems, we propose a multi‐scale framework that combines detailed materials characterization, optimization of plastic‐biomolecular interfacial interactions, and enhancement of biocatalytic kinetics to develop effective plastic‐deconstructing enzymes. This approach leverages principles from reaction kinetics, transport and interfacial phenomena, and enzyme engineering to systematically address barriers across diverse plastic types. Our framework aims to accelerate the discovery and optimization of biocatalysts capable of scalable, selective, and efficient deconstruction of plastic waste. These advances hold potential to enable sustainable biological recycling and upcycling pathways, contributing to global efforts in mitigating plastic pollution and promoting circular material economies.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗