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DOE OSTI2022

Heterotropic roles of divalent cations in the establishment of allostery and affinity maturation of integrin αXβ2

Allosteric activation and silencing of leukocyte β2-integrins transpire through cation-dependent structural changes, which mediate integrin biosynthesis and recycling, and are essential to designing leukocyte-specific drugs. Stepwise addition of Mg 2+ reveals two mutually coupled events for the αXβ2 ligand-binding domain—the αX I-domain—corresponding to allostery establishment and affinity maturation. Electrostatic alterations in the Mg 2+ -binding site establish long-range couplings, leading to both pH- and Mg 2+ -occupancy-dependent biphasic stability change in the αX I-domain fold. The ligand-binding sensorgrams show composite affinity events for the αX I-domain accounting for the multiplicity of the αX I-domain conformational states existing in the solution. On cell surfaces, increasing Mg 2+ concentration enhanced adhesiveness of αXβ2. This work highlights how intrinsically flexible pH- and cation-sensitive architecture endows a unique dynamic continuum to the αI-domain structure on the intact integrin, thereby revealing the importance of allostery establishment and affinity maturation in both extracellular and intracellular integrin events.

36 MATERIALS SCIENCE↗

DOE OSTI2023

Artificial extracellular matrix scaffolds of mobile molecules enhance maturation of human stem cell-derived neurons

Human induced pluripotent stem cell (hiPSC) technologies offer a unique resource for modeling neurological diseases. However, iPSC models are fraught with technical limitations including abnormal aggregation and inefficient maturation of differentiated neurons. These problems are in part due to the absence of synergistic cues of the native extracellular matrix (ECM). We report on the use of three artificial ECMs based on peptide amphiphile (PA) supramolecular nanofibers. All nanofibers display the laminin-derived IKVAV signal on their surface but differ in the nature of their non-bioactive domains. Here, we find that nanofibers with greater intensity of internal supramolecular motion have enhanced bioactivity toward hiPSC-derived motor and cortical neurons. Proteomic, biochemical, and functional assays reveal that highly mobile PA scaffolds caused enhanced β1-integrin pathway activation, reduced aggregation, increased arborization, and matured electrophysiological activity of neurons. Our work highlights the importance of designing biomimetic ECMs to study the development, function, and dysfunction of human neurons.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2025

Unravelling the dynamics of the maturation protein in MS2 bacteriophage via molecular simulations

The MS2 bacteriophage capsid serves as a model system for studying viral structure and function. Mature MS2 virus consists of 178 capsid proteins and a single maturation protein (MP), which is essential for host receptor binding and infection initiation. Despite its critical role, the dynamic behavior of the capsid with the MP remains poorly understood. To address this, we conducted 0.5 µs all-atom molecular dynamics (MD) simulations of the MS2 capsid with and without the MP, revealing key insights into its structural dynamics. Our simulations showed that MP exhibits high flexibility, particularly in the “tip” and “side-loop” regions, which undergo significant motions that likely enhance its ability to engage with the F-pilus receptor. Detailed analysis of MP conformational states revealed that loop rearrangements around H357 enable transient switching between “semi-closed” and “open” conformations, suggesting a conformational selection mechanism for pilus binding. Additionally, ion interaction analyses revealed distinct sodium and chloride binding patterns, where sodium ions were mostly found at the outer capsid shell, while chloride ions interacted with the basic residues on the RNA-facing side. We also found that the presence of the MP enhances salt-bridge interactions, contributing to increased capsid stability, yet it does not significantly alter the pore sizes of pentameric and hexameric units. Together, these findings provide new insights into the functional role of the MP, highlighting its contribution to capsid stability and host receptor engagement. This study offers a foundation for understanding capsid dynamics relevant to viral infectivity and may guide future rational strategies aimed at disrupting host-virus interactions.

Capsid stability↗

NASA NTRS2022

TESS Hunt for Young and Maturing Exoplanets (THYME). VI. An 11 Myr Giant Planet Transiting a Very-low-mass Star in Lower Centaurus Crux

Mature super-Earths and sub-Neptunes are predicted to be ≃ Jovian radius when younger than 10 Myr. Thus, we expect to find 5-15 R ⊕ planets around young stars even if their older counterparts harbor none. We report the discovery and validation of TOI 1227b, a 0.85 ± 0.05 R J (9.5 R ⊕) planet transiting a very-low-mass star (0.170 ± 0.015 M ⊙) every 27.4 days. TOI 1227's kinematics and strong lithium absorption confirm that it is a member of a previously discovered subgroup in the Lower Centaurus Crux OB association, which we designate the Musca group. We derive an age of 11 ± 2 Myr for Musca, based on lithium, rotation, and the color-magnitude diagram of Musca members. The TESS data and ground-based follow-up show a deep (2.5%) transit. We use multiwavelength transit observations and radial velocities from the IGRINS spectrograph to validate the signal as planetary in nature, and we obtain an upper limit on the planet mass of ≃0.5 M J. Because such large planets are exceptionally rare around mature low-mass stars, we suggest that TOI 1227b is still contracting and will eventually turn into one of the more common <5 R ⊕ planets.

5–15 R⊕ planets↗

DOE OSTI2023

Insights into the mechanism of SARS-CoV-2 main protease autocatalytic maturation from model precursors

A critical step for SARS-CoV-2 assembly and maturation involves the autoactivation of the main protease (MPro WT ) from precursor polyproteins. Upon expression, a model precursor of MPro WT mediates its own release at its termini rapidly to yield a mature dimer. A construct with an E290A mutation within MPro exhibits time dependent autoprocessing of the accumulated precursor at the N-terminal nsp4/nsp5 site followed by the C-terminal nsp5/nsp6 cleavage. In contrast, a precursor containing E290A and R298A mutations (MPro M ) displays cleavage only at the nsp4/nsp5 site to yield an intermediate monomeric product, which is cleaved at the nsp5/nsp6 site only by MPro WT . MPro M and the catalytic domain (MPro 1-199 ) fused to the truncated nsp4 region also show time-dependent conversion in vitro to produce MPro M and MPro 1-199 , respectively. The reactions follow first-order kinetics indicating that the nsp4/nsp5 cleavage occurs via an intramolecular mechanism. These results support a mechanism involving an N-terminal intramolecular cleavage leading to an increase in the dimer population and followed by an intermolecular cleavage at the C-terminus. Thus, targeting the predominantly monomeric MPro precursor for inhibition may lead to the identification of potent drugs for treatment.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

From genes to function: regulation, maturation, and evolution of cytochrome c nitrite reductase in nitrate reduction to ammonium

ABSTRACT The dissimilatory nitrate reduction to ammonium pathway converts nitrate to ammonium, a vital reaction in the global nitrogen cycle. The second step of the pathway is performed by cytochromecnitrite reductase (NrfA), a soluble, periplasmic cytochrome responsible for the reduction of nitrite to ammonium. The pentaheme NrfA catalyzes this six-electron and eight-proton reduction of nitrite at a single active site with the help of its quinol oxidase partners. In this review, we discuss our current understanding of (i) the structure, homology, and evolution of both NrfA and its redox partners, (ii) the regulation of thenrfoperon, and (iii) the maturation of NrfA proteins via unique cytochrome maturation pathways.

Biotechnology & Applied Microbiology↗

DOE OSTI2024

Survival and Age at Maturity in Head-Started Wood Turtles ( Glyptemys insculpta ) with Implications for Population Recovery

A small relict population of Glyptemys insculpta (Wood Turtle) was discovered on a protected area in New Jersey in 2006. Marking and radio-tracking of the old founder individuals helped to determine movement patterns and habitat use. Monitoring of nesting females revealed that nesting habitat and nest success was limited due to invasive plants, human landscape alteration, and Procyon lotor (Raccoon) depredation. We initiated habitat restoration including creation of protected nesting areas, mowing in winter, invasive plant removal, and adjacent landowner education. Here, we direct-released hatchlings from protected nests from 2006 to 2015, yet only a few were detected in subsequent years. Between 2011 and 2023, some or all of each hatchling cohort were head-started indoors at a high school for 9 months. We have continuously radio-tracked all head-starts from the 2011 cohort and portions of the 2012–2014 cohorts. Head-started turtles found their own food, established home ranges, and hibernated communally with founder adults. Subsidized Raccoons, mowers, automobiles, and flooding events—all human-instigated—were the causes of mortality. The first males and first females from the 2011 head-start cohort reached maturity in 2017 and 2019, respectively, at ages 6–8, younger than expected by 4–5 years. Successful reproduction by head-starts was confirmed by viable hatchlings produced in 2019, 2020, and 2023. Head-starting can pull a relict population out of the nose-dive to extirpation when used in conjunction with habitat-restoration practices, but it must be conducted with persistence and continuity over at least the number of years it takes for the earliest cohorts to reach maturity and begin producing offspring of their own.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

AmeriFlux FLUXNET-1F CA-TPD Ontario - Turkey Point Mature Deciduous

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site CA-TPD Ontario - Turkey Point Mature Deciduous. This is the FLUXNET version of the carbon flux data for the site CA-TPD Ontario - Turkey Point Mature Deciduous produced by applying the standard ONEFlux (1F) software. Site Description - The forest is approximately 90 years old. Naturally regenerated on sandy terrain and abandoned agricultural land. Predominantly hardwood species with a few scattered conifers. Site has been managed (thinned) in the past. It has a high biodiversity with 573 tree and plant species, 102 bird species, 23 mamal species and 22 reptile and amphibian species (SWALSREP Report, 1999). The dominant tree species is white oak (Quercus alba), with other scattered broadleaf Carolinian species including sugar and red maple (Acer saccharum, A. rubrum), American beech (Fagus grandifolia), black and red oak (Q. velutina, Q. rubra) and white ash (Fraxinus americana) . There are also scattered conifers, mostly white and red pine (Pinus strobes, P. resinosa), comprising about 5% of the trees. Average tree height is 25.7 m with a stand density of 504 ± 18 trees per hectare. Average tree diameter at breast height is 22.3 cm and basal area is 0.06 m2 or approximately 29 square meters per hectare.

Arain, M. Altaf↗

DOE OSTI2023

AmeriFlux FLUXNET-1F CA-Obs Saskatchewan - Western Boreal, Mature Black Spruce

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site CA-Obs Saskatchewan - Western Boreal, Mature Black Spruce. This is the FLUXNET version of the carbon flux data for the site CA-Obs Saskatchewan - Western Boreal, Mature Black Spruce produced by applying the standard ONEFlux (1F) software. Site Description - 53.98717° N, 105.11779° W, elavation of 628.94 m, BOREAS 1994, 1996, BERMS climate measurements began Dec. 1996 and flux measurements in Apr. 1999

Black, T. Andrew↗

DOE OSTI2023

AmeriFlux FLUXNET-1F CA-Qfo Quebec - Eastern Boreal, Mature Black Spruce

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site CA-Qfo Quebec - Eastern Boreal, Mature Black Spruce. This is the FLUXNET version of the carbon flux data for the site CA-Qfo Quebec - Eastern Boreal, Mature Black Spruce produced by applying the standard ONEFlux (1F) software. Site Description - 49.69247° N / 74.34204° W, elevation of 387 mm, 90 - 100 yr old Black Spruce, Jack Pine, feather moss

Margolis, Hank A.↗

DOE OSTI2026

AmeriFlux FLUXNET-1F CA-Na1 New Brunswick - 1967 Balsam Fir - Nashwaak Lake Site 01 (Mature balsam fir forest)

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site CA-Na1 New Brunswick - 1967 Balsam Fir - Nashwaak Lake Site 01 (Mature balsam fir forest). This is the FLUXNET version of the carbon flux data for the site CA-Na1 New Brunswick - 1967 Balsam Fir - Nashwaak Lake Site 01 (Mature balsam fir forest) produced by applying the standard ONEFlux (1F) software. Site Description - immature balsam fir forest

Bourque, Charles P.-A.↗

DOE OSTI2026

AmeriFlux FLUXNET-1F CA-Ojp Saskatchewan - Western Boreal, Mature Jack Pine

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site CA-Ojp Saskatchewan - Western Boreal, Mature Jack Pine. This is the FLUXNET version of the carbon flux data for the site CA-Ojp Saskatchewan - Western Boreal, Mature Jack Pine produced by applying the standard ONEFlux (1F) software. Site Description - 53.91634° N, 104.69203° W, elavation of 579.27 m, BOREAS 1994, BERMS climate measurements began Mar. 1997 and flux measurements Aug. 1999

Black, Andrew T. [University of British Columbia]↗

DOE OSTI2024

SHASTA: Task 2.2: Sensor Development for Subsurface Hydrogen Storage Monitoring Technology Maturation and Commercialization Plan (TMCP)

In Task 2.2 under the SHASTA program, NETL is developing in-situ optical fiber sensors for real-time monitoring of hydrogen, methane, and chemical parameters such as pH at subsurface hydrogen storage conditions. This Technology Maturation and Commercialization Plan report will provide the summary of NETL sensor development and test results under the SHASTA program to meet the subsurface application requirements and proposed technology maturation and deployment plan, and commercialization potential.

08 HYDROGEN↗

DOE OSTI2025

Moving Beyond Direct Load Control: A Maturity Model for Realizing the Promise of Demand Flexibility

This report discusses a new maturity model that regulators and utilities can use to guide and expand demand flexibility programs and enable the resources to provide more grid services. The model has six demand flexibility categories: planning and design; customer engagement; program operations; evaluation, measurement and verification; distributed energy resource orchestration; and data infrastructure. Within each category, capabilities are identified and described on a maturity scale that ranges from performing below expectations to improving on best practices.

24 POWER TRANSMISSION AND DISTRIBUTION↗

NASA NTRS1975

Remotely sensing wheat maturation with radar

The scattering properties of wheat were studied in the 8-18 GHz band as a function of frequency, polarization, incidence angle, and crop maturity. Supporting ground truth was collected at the time of measurement. The data indicate that the radar backscattering coefficient is sensitive to both radar system parameters and crop characteristics particularly at incidence angles near nadir. Linear regression analyses of the radar backscattering coefficient on both time and plant moisture content result in rather good correlation. Furthermore, by calculating the average time rate of change of the radar backscattering coefficient it is found that it undergoes rapid variations shortly before and after the wheat is harvested. Both of these analyses suggest methods for estimating wheat maturity and for monitoring the progress of harvest.

Bush, T. F.↗

NASA NTRS1975

Effects of maturation on the reflectance of the lunar regolith: Apollo 16 - A case study

Soils at the Apollo 16 site become progressively darker as the percentage of glassy agglutinates increases. Magnetic separates of the agglutinate fraction of a soil are always darker than the bulk soil and are darker than the nonagglutinate fraction consisting of rock and mineral fragments. Darkening of a soil with maturity is due mainly to the increasing proportion of agglutinates. Coating of rock and mineral fragments with thin deposits of glass aids darkening in a minor way, but most of these particles eventually are destroyed by melting as the soils mature.

Adams, J. B.↗

NASA NTRS1977

Soil maturity and planetary regoliths - The Moon, Mercury, and the asteroids

Observations of the Moon, Mercury and the asteroids provide data which can be used to assess current ideas about the optical maturation of regoliths of bodies without atmospheres. Telescopic data show Mercury's surface to be as optically mature as the Moon's while the asteroids appear to have 'fresh' surfaces. We suggest that these cases illustrate two different paths of regolith evolution caused by differing micrometeoroid impact velocities, gravity, and in some cases composition.

Matson, D. L.↗

NASA NTRS1978

Primordial Pb, radiogenic Pb and lunar soil maturation

Pb-204 is directly correlated with the reduced Fe measured by ferromagnetic resonance. A similar correlation has been noted for hydrolyzable carbon (Pillinger et al., 1974). An enrichment of these elements appears to have occurred during soil maturation. In contrast to Pb-204, radiogenic Pb is reported to be lost during soil maturation (Church et al., 1976). Radiogenic Pb is present in mineral grains and may be lost by solar wind sputtering (or volatilization) and not resupplied. Pb-204 coating grain surfaces acts as a reservoir to provide the Pb-204 being extracted in the reduced Fe formation process. Venting or some other volatile release mechanism may replenish the surface-related Pb-204.

Reed, G. W., Jr.↗