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Flavin disulfide reductases (FDRs) are FAD-dependent enzymes that transmit electrons from NAD(P)H to reduce specific oxidant substrate disulfides. These enzymes have been studied extensively, most particularly the paradigm examples: glutathione reductase and thioredoxin reductase. The common, though not universal, traits of the family include a tyrosine- or phenylalanine-gated binding pocket for NAD(P) nicotinamides adjacent to the FAD isoalloxazine re-face, and a disulfide stacked against the si-face of the isoalloxazine whose dithiol form is activated for subsequent exchange reactions by a nearby histidine acting as a base. This arrangement promotes transduction of the reducing equivalents for disulfide exchange relay reactions. From an observational standpoint the proximal parallel stacking of three redox moieties induces up to three opportunities for unique charge transfer interactions (NAD(P)H FAD, NAD(P)+•FADH2, and FAD•thiolate). In transient state, the charge transfer transitions provide discrete signals to assign reaction sequences. This review summarizes the lineage of observations for the FDR enzymes that have been extensively studied. Where applicable and in order to chart a consistent interpretation of the record, only data derived from studies that used anaerobic methods are cited. These data reveal a recurring theme for catalysis that is elaborated with specific additional functionalities for each oxidant substrate. 

Seafloor moorings measuring pressure and temperature were deployed from April to September 2023 at three sites near the route of the fiber optic telecommunications cable that extends offshore of Oliktok Point, Alaska. The raw data data (1 Hertz (Hz) sampling) are processed for hourly estimates of the ocean surface wave conditions, along with average seawater temperature and average depth. The sites were ice-covered from April to July, then mostly open water in August and September. The data were collected to calibrate proxy wave measurements using Distributed Acoustic Sensing (DAS) from the telecommunications cable. 

Thioredoxin/glutathione reductase (TGR) from the platyhelminthic parasitic worms has recently been identified as a drug target for the treatment of schistosomiasis. Schistosomes lack catalase, and so are heavily reliant on the regeneration of reduced thioredoxin (Trx) and glutathione (GSH) to reduce peroxiredoxins that ameliorate oxidative damage from hydrogen peroxide generated by the host immune response. This study focuses on the characterization of the catalytic mechanism ofSchistosoma mansoniTGR (SmTGR). Variant forms of SmTGR were studied to assign the function of residues that participate in the electron distribution chain within the enzyme. Using anaerobic transient state spectrophotometric methods, redox changes for the FAD and NADPH were observed and the function of specific residues was defined from observation of charge transfer absorption transitions that are indicative of specific complexations and redox states. The C159S variant prevented distribution of electrons beyond the flavin and as such did not accumulate thiolate-FAD charge transfer absorption. The lack of this absorption facilitated observation of a new charge transfer absorption consistent with proximity of NADPH and FAD. The C159S variant was used to confine electrons from NADPH at the flavin, and it was shown that NADPH and FAD exchange hydride in both directions and come to an equilibrium that yields only fractional FAD reduction, suggesting that both have similar reduction potentials. Mutation of U597 to serine resulted in sustained thiolate-FAD charge transfer absorption and loss of the ability to reduce Trx, indicating that the C596-U597 disulfide functions in the catalytic sequence to receive electrons from the C154 C159 pair and distribute them to Trx. No kinetic evidence for a loss or change in function associated with the distal C28-C31 disulfide was observed when the C31S variant reductive half-reaction was observed. The Y296A variant was shown to slow the rate of but increase extent of reduction of the flavin, and the dissociation of NADP⁺. The H571 residue was confirmed to be the residue responsible for the deprotonation of the C159 thiol, increasing its reactivity and generating the prominent thiolate-FAD charge transfer absorption that accumulates with oxidation of the flavin.

 

We present results from wide-field imaging of the resolved stellar populations of the dwarf spheroidal galaxies Cassiopeia III (And XXXII) and Perseus I (And XXXIII), two satellites in the outer stellar halo of the Andromeda galaxy (M31). Our WIYN pODI photometry traces the red giant star population in each galaxy to ∼2.5−3 half-light radii from the galaxy center. We use the tip of the red giant branch (TRGB) method to derive distances of (m−M)₀= 24.62 ± 0.12 mag (839${}_{-45}^{+48}$kpc, or${156}_{-13}^{+16}$kpc from M31) for Cas III and 24.47 ± 0.13 mag (738${}_{-45}^{+48}$kpc, or 351${}_{-16}^{+17}$kpc from M31) for Per I. These values are consistent within the errors with TRGB distances derived from a deeper Hubble Space Telescope study of the galaxies’ inner regions. For each galaxy, we derive structural parameters, total magnitude, and central surface brightness. We also place upper limits on the ratio of neutral hydrogen gas mass to optical luminosity, confirming the gas-poor nature of both galaxies. We combine our data set with corresponding data for the M31 satellite galaxy Lacerta I (And XXXI) from earlier work and search for substructure within the RGB star populations of Cas III, Per I, and Lac I. We find an overdense region on the west side of Lac I at a significance level of 2.5σ–3σand a low-significance filament extending in the direction of M31. In Cas III, we identify two modestly significant overdensities near the center of the galaxy and another at two half-light radii. Per I shows no evidence for substructure in its RGB star population, which may reflect this galaxy’s isolated nature.

 

Thioredoxin/glutathione reductase from Schistosoma mansoni (SmTGR) catalyzes the reduction of both oxidized thioredoxin and glutathione with electrons from reduced nicotinamide adenine dinucleotide phosphate (NADPH). SmTGR is a drug target for the treatment of Schistosomiasis, an infection caused by Schistosoma platyhelminths residing in the blood vessels of the host. Schistosoma spp. are reliant on TGR enzymes as they lack catalase and so use reduced thioredoxin and glutathione to regenerate peroxiredoxins consumed in the detoxification of reactive oxygen species. SmTGR is a flavin adenine dinucleotide (FAD)-dependent enzyme, and we have used the flavin as a spectrophotometric reporter to observe the movement of electrons within the enzyme. The data show that NADPH fractionally reduces the active site flavin with an observed rate constant estimated in this study to be ∼3000 s-1. The flavin then reoxidizes by passing electrons at a similar rate to the proximal Cys159-Cys154 disulfide pair. The dissociation of NADP+ occurs with a rate of ∼180 s-1, which induces the deprotonation of Cys159, and this coincides with the accumulation of an intense FAD-thiolate charge transfer band. It is proposed that the electrons then pass to the Cys596-Cys597 disulfide pair of the associated subunit in the dimer with a net rate constant of ∼2 s-1. (Note: Cys597 is Sec597 in wild-type (WT) SmTGR.) From this position, the electrons can be passed to oxidized thioredoxin or further into the protein to reduce the Cys28-Cys31 disulfide pair of the originating subunit of the dimer. From the Cys28-Cys31 center, electrons can then pass to oxidized glutathione that has a binding site directly adjacent. 

Dihydropyrimidine dehydrogenase (DPD) is a flavin dependent enzyme that catalyzes the reduction of the 5,6-vinylic bond of pyrimidines uracil and thymine with electrons from NADPH. DPD has two active sites that are separated by ∼60 Å. At one site NADPH binds adjacent to an FAD cofactor and at the other pyrimidine binds proximal to an FMN. Four Fe4S4 centers span the distance between these active sites. It has recently been established that the enzyme undergoes reductive activation prior to reducing the pyrimidine. In this initial process NADPH is oxidized at the FAD site and electrons are transmitted to the FMN via the Fe4S4 centers to yield the active state with a cofactor set of FAD•4(Fe4S4)•FMNH2. The catalytic chemistry of DPD can be studied in transient-state by observation of either NADPH consumption or charge transfer absorption associated with complexation of NADPH adjacent to the FAD. Here we have utilized both sets of absorption transitions to find evidence for specific additional aspects of the DPD mechanism. Competition for binding with NADP+ indicates that the two charge transfer species observed in activation/single turnover reactions arise from NADPH populating the FAD site before and after reductive activation. An additional charge transfer species is observed to accumulate at longer times when high NADPH concentrations are mixed with the enzyme•pyrimidine complex and this data can be modelled based on asymmetry in the homodimer. It was also shown that, like pyrimidines, dihydropyrimidines induce rapid reductive activation indicating that the reduced pyrimidine formed in turnover can stimulate the reinstatement of the active state of the enzyme. Investigation of the reverse reaction revealed that dihydropyrimidines alone can reductively activate the enzyme, albeit inefficiently. In the presence of dihydropyrimidine and NADP+ DPD will form NADPH but apparently without measurable reductive activation. Pyrimidines that have 5-substituent halogens were utilized to probe both reductive activation and turnover. The linearity of the Hammett plot based on the rate of hydride transfer to the pyrimidine establishes that, at least to the radius of an iodo-group, the 5-substituent volume does not have influence on the observed kinetics of pyrimidine reduction. 

The microstructure of the uppermost portions of a melting Arctic sea ice cover has a disproportionately large influence on how incident sunlight is reflected and absorbed in the ice/ocean system. The surface scattering layer (SSL) effectively backscatters solar radiation and keeps the surface albedo of melting ice relatively high compared to ice with the SSL manually removed. Measurements of albedo provide information on how incoming shortwave radiation is partitioned by the SSL and have been pivotal to improving climate model parameterizations. However, the relationship between the physical and optical properties of the SSL is still poorly constrained. Until now, radiative transfer models have been the only way to infer the microstructure of the SSL. During the MOSAiC expedition of 2019–2020, we took samples and, for the first time, directly measured the microstructure of the SSL on bare sea ice using X-ray micro-computed tomography. We show that the SSL has a highly anisotropic, coarse, and porous structure, with a small optical diameter and density at the surface, increasing with depth. As the melting surface ablates, the SSL regenerates, maintaining some aspects of its microstructure throughout the melt season. We used the microstructure measurements with a radiative transfer model to improve our understanding of the relationship between physical properties and optical properties at 850 nm wavelength. When the microstructure is used as model input, we see a 10–15% overestimation of the reflectance at 850 nm. This comparison suggests that either a) spatial variability at the meter scale is important for the two in situ optical measurements and therefore a larger sample size is needed to represent the microstructure or b) future work should investigate either i) using a ray-tracing approach instead of explicitly solving the radiative transfer equation or ii) using a more appropriate radiative transfer model.

 

Snow plays an essential role in the Arctic as the interface between the sea ice and the atmosphere. Optical properties, thermal conductivity and mass distribution are critical to understanding the complex Arctic sea ice system’s energy balance and mass distribution. By conducting measurements from October 2019 to September 2020 on the Multidisciplinary drifting Observatory for the Study of Arctic Climate (MOSAiC) expedition, we have produced a dataset capturing the year-long evolution of the physical properties of the snow and surface scattering layer, a highly porous surface layer on Arctic sea ice that evolves due to preferential melt at the ice grain boundaries. The dataset includes measurements of snow during MOSAiC. Measurements included profiles of depth, density, temperature, snow water equivalent, penetration resistance, stable water isotope, salinity and microcomputer tomography samples. Most snowpit sites were visited and measured weekly to capture the temporal evolution of the physical properties of snow. The compiled dataset includes 576 snowpits and describes snow conditions during the MOSAiC expedition.

 

Abstract. The melting of sea ice floes from the edges (lateral melting) results in open-water formation and subsequently increases absorption of solar shortwave energy. However, lateral melt plays a small role in the sea ice mass budget in both hemispheres in most climate models. This is likely influenced by the simple parameterization of lateral melting in sea ice models that are constrained by limited observations. Here we use a coupled climate model (CESM2.0) to assess the sensitivity of modeled sea ice state to the lateral melt parameterization in preindustrial and 2×CO2 runs. The runs explore the implications of how lateral melting is parameterized and structural changes in how it is applied. The results show that sea ice is sensitive both to the parameters determining the effective lateral melt rate and the nuances in how lateral melting is applied to the ice pack. Increasing the lateral melt rate is largely compensated for by decreases in the basal melt rate but still results in a significant decrease in sea ice concentration and thickness, particularly in the marginal ice zone. Our analysis suggests that this is tied to the increased efficiency of lateral melting at forming open water during the summer melt season, which drives the majority of the ice–albedo feedback. The more seasonal Southern Hemisphere ice cover undergoes larger relative reductions in sea ice concentration and thickness for the same relative increase in lateral melt rate, likely due to the hemispheric differences in the role of the sea-ice–upper-ocean coupling.Additionally, increasing the lateral melt rate under a 2×CO2 forcing, where sea ice is thinner, results in a smaller relative change in sea ice mean state but suggests that open-water-formation feedbacks are likely to steepen the decline to ice-free summer conditions.Overall, melt processes are more efficient at forming open water in thinner ice scenarios (as we are likely to see in the future), suggesting the importance of accurately representing thermodynamic evolution. Revisiting model parameterizations of lateral melting with observations will require finding new ways to represent salient physical processes. 

Leads play an important role in the exchange of heat, gases, vapour, and particles between seawater and the atmosphere in ice-covered polar oceans. In summer, these processes can be modified significantly by the formation of a meltwater layer at the surface, yet we know little about the dynamics of meltwater layer formation and persistence. During the drift campaign of the Multidisciplinary drifting Observatory for the Study of Arctic Climate (MOSAiC), we examined how variation in lead width, re-freezing, and mixing events affected the vertical structure of lead waters during late summer in the central Arctic. At the beginning of the 4-week survey period, a meltwater layer occupied the surface 0.8 m of the lead, and temperature and salinity showed strong vertical gradients. Stable oxygen isotopes indicate that the meltwater consisted mainly of sea ice meltwater rather than snow meltwater. During the first half of the survey period (before freezing), the meltwater layer thickness decreased rapidly as lead width increased and stretched the layer horizontally. During the latter half of the survey period (after freezing of the lead surface), stratification weakened and the meltwater layer became thinner before disappearing completely due to surface ice formation and mixing processes. Removal of meltwater during surface ice formation explained about 43% of the reduction in thickness of the meltwater layer. The remaining approximate 57% could be explained by mixing within the water column initiated by disturbance of the lower boundary of the meltwater layer through wind-induced ice floe drift. These results indicate that rapid, dynamic changes to lead water structure can have potentially significant effects on the exchange of physical and biogeochemical components throughout the atmosphere–lead–underlying seawater system.