Enzymes must be ordered to allow the stabilization of transition states by their active sites, yet dynamic enough to adopt alternative conformations suited to other steps in their catalytic cycles. ...The biophysical principles that determine how specific protein dynamics evolve and how remote mutations affect catalytic activity are poorly understood. Here we examine a 'molecular fossil record' that was recently obtained during the laboratory evolution of a phosphotriesterase from Pseudomonas diminuta to an arylesterase. Analysis of the structures and dynamics of nine protein variants along this trajectory, and three rationally designed variants, reveals cycles of structural destabilization and repair, evolutionary pressure to 'freeze out' unproductive motions and sampling of distinct conformations with specific catalytic properties in bi-functional intermediates. This work establishes that changes to the conformational landscapes of proteins are an essential aspect of molecular evolution and that change in function can be achieved through enrichment of preexisting conformational sub-states.
•Laboratory directed evolution and ancestral reconstruction can facilitate study of protein structural dynamics.•Advances in biophysical and computational techniques facilitate this ...investigation.•Conformational dynamism is valuable in the acquisition of new function, as multiple potentially productive conformations can be sampled.•Evolution ‘freezes out’ unproductive motions, reducing unnecessary dynamics.
This review focuses on recent work that has begun to establish specific functional roles for protein conformational dynamics, specifically how the conformational landscapes that proteins can sample can evolve under laboratory based evolutionary selection. We discuss recent technical advances in computational and biophysical chemistry, which have provided us with new ways to dissect evolutionary processes. Finally, we offer some perspectives on the emerging view of conformational dynamics and evolution, and the challenges that we face in rationally engineering conformational dynamics.
Abstract
Serine protease inhibitors (serpins) are found in all kingdoms of life and play essential roles in multiple physiological processes. Owing to the diversity of the superfamily, phylogenetic ...analysis is challenging and prokaryotic serpins have been speculated to have been acquired from Metazoa through horizontal gene transfer due to their unexpectedly high homology. Here, we have leveraged a structural alignment of diverse serpins to generate a comprehensive 6,000-sequence phylogeny that encompasses serpins from all kingdoms of life. We show that in addition to a central “hub” of highly conserved serpins, there has been extensive diversification of the superfamily into many novel functional clades. Our analysis indicates that the hub proteins are ancient and are similar because of convergent evolution, rather than the alternative hypothesis of horizontal gene transfer. This work clarifies longstanding questions in the evolution of serpins and provides new directions for research in the field of serpin biology.
Some of the greatest transformations in vertebrate history involve developmental and evolutionary origins of avian flight. Flight is the most power-demanding mode of locomotion, and volant adult ...birds have many anatomical features that presumably help meet these demands. However, juvenile birds, like the first winged dinosaurs, lack many hallmarks of advanced flight capacity. Instead of large wings they have small "protowings", and instead of robust, interlocking forelimb skeletons their limbs are more gracile and their joints less constrained. Such traits are often thought to preclude extinct theropods from powered flight, yet young birds with similarly rudimentary anatomies flap-run up slopes and even briefly fly, thereby challenging longstanding ideas on skeletal and feather function in the theropod-avian lineage. Though skeletons and feathers are the common link between extinct and extant theropods and figure prominently in discussions on flight performance (extant birds) and flight origins (extinct theropods), skeletal inter-workings are hidden from view and their functional relationship with aerodynamically active wings is not known. For the first time, we use X-ray Reconstruction of Moving Morphology to visualize skeletal movement in developing birds, and explore how development of the avian flight apparatus corresponds with ontogenetic trajectories in skeletal kinematics, aerodynamic performance, and the locomotor transition from pre-flight flapping behaviors to full flight capacity. Our findings reveal that developing chukars (Alectoris chukar) with rudimentary flight apparatuses acquire an "avian" flight stroke early in ontogeny, initially by using their wings and legs cooperatively and, as they acquire flight capacity, counteracting ontogenetic increases in aerodynamic output with greater skeletal channelization. In conjunction with previous work, juvenile birds thereby demonstrate that the initial function of developing wings is to enhance leg performance, and that aerodynamically active, flapping wings might better be viewed as adaptations or exaptations for enhancing leg performance.
Abstract
Groundwater discharge to streams is a nonpoint source of nitrogen (N) that confounds N mitigation efforts and represents a significant portion of the annual N loading to watersheds. However, ...we lack an understanding of where and how much groundwater N enters streams and watersheds. Nitrogen concentrations at the end of groundwater flowpaths are the culmination of biogeochemical and physical processes from the contributing land area where groundwater recharges, within the aquifer system, and in the near-stream riparian area where groundwater discharges to streams. Our research objectives were to quantify the spatial distribution of N concentrations at groundwater discharges throughout a mixed land-use watershed and to evaluate how relationships among contributing and riparian land cover, modeled aquifer characteristics, and groundwater discharge biogeochemistry explain the spatial variation in groundwater discharge N concentrations. We accomplished this by integrating high-resolution thermal infrared surveys to locate groundwater discharge, biogeochemical sampling of groundwater, and a particle tracking model that links groundwater discharge locations to their contributing area land cover. Groundwater N loading from groundwater discharges within the watershed varied substantially between and within streambank groundwater discharge features. Groundwater nitrate concentrations were spatially heterogeneous ranging from below 0.03–11.45 mg-N/L, varying up to 20-fold within meters. When combined with the particle tracking model results and land cover metrics, we found that groundwater discharge nitrate concentrations were best predicted by a linear mixed-effect model that explained over 60% of the variation in nitrate concentrations, including aquifer chemistry (dissolved oxygen, Cl
−
, SO
4
2−
), riparian area forested land cover, and modeled physical aquifer characteristics (discharge, Euclidean distance). Our work highlights the significant spatial variability in groundwater discharge nitrate concentrations within mixed land-use watersheds and the need to understand groundwater N processing across the many spatiotemporal scales within groundwater cycling.
Monoclonal antibodies derived from blood plasma cells of acute HIV-1-infected individuals are predominantly targeted to the HIV Env gp41 and cross-reactive with commensal bacteria. To understand this ...phenomenon, we examined anti-HIV responses in ileum B cells using recombinant antibody technology and probed their relationship to commensal bacteria. The dominant ileum B cell response was to Env gp41. Remarkably, a majority (82%) of the ileum anti-gp41 antibodies cross-reacted with commensal bacteria, and of those, 43% showed non-HIV-1 antigen polyreactivity. Pyrosequencing revealed shared HIV-1 antibody clonal lineages between ileum and blood. Mutated immunoglobulin G antibodies cross-reactive with both Env gp41 and microbiota could also be isolated from the ileum of HIV-1 uninfected individuals. Thus, the gp41 commensal bacterial antigen cross-reactive antibodies originate in the intestine, and the gp41 Env response in HIV-1 infection can be derived from a preinfection memory B cell pool triggered by commensal bacteria that cross-react with Env.
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•HIV-1-reactive B cells in the terminal ileum primarily target Env gp41•A majority of the HIV-1 gp41-reactive antibodies cross- react with commensal bacteria•B cell clonal lineages are shared between the terminal ileum and blood•Env gp41 and microbiota cross-reactive IgG can be isolated from HIV-1 naive individuals
The initial, nonneutralizing antibody response to HIV-1 may derive from memory B cells cross-reactive with environmental antigens. By isolating antibodies from ileum B cells, Trama et al. provide evidence that a preinfection pool of microbiota-reactive B cells can be the origin of the antibody response to acute HIV-1 infection.
Previous studies of severe acute respiratory syndrome coronavirus 2 infection in infants have incompletely characterized factors associated with severe illness or focused on infants born to mothers ...with coronavirus disease 2019 (COVID-19). Here we highlight demographics, clinical characteristics and laboratory values that differ between infants with and without severe acute COVID-19.
Active surveillance was performed by the Overcoming COVID-19 network to identify children and adolescents with severe acute respiratory syndrome coronavirus 2-related illness hospitalized at 62 sites in 31 states from March 15 to December 27, 2020. We analyzed patients >7 days to <1 year old hospitalized with symptomatic acute COVID-19.
We report 232 infants >7 days to <1 year of age hospitalized with acute symptomatic COVID-19 from 37 US hospitals in our cohort from March 15 to December 27, 2020. Among 630 cases of severe COVID-19 in patients >7 days to <18 years old, 128 (20.3%) were infants. In infants with severe illness from the entire study period, the median age was 2 months, 66% were from racial and ethnic minority groups, 66% were previously healthy, 73% had respiratory complications, 13% received mechanical ventilation and <1% died.
Infants accounted for over a fifth of children <18 years of age hospitalized for severe acute COVID-19, commonly manifesting with respiratory symptoms and complications. Although most infants hospitalized with COVID-19 did not suffer significant complications, longer term outcomes remain unclear. Notably, 75% of infants with severe disease were <6 months of age in this cohort study period, which predated maternal COVID-19 vaccination, underscoring the importance of maternal vaccination for COVID-19 in protecting the mother and infant.
Abstract Preferential groundwater discharge features along stream corridors are ecologically important at local and stream network scales, yet we lack quantification of the multiscale controls on the ...spatial patterning of groundwater discharge. Here we identify physical attributes that best explain variation in the presence and lateral extent of preferential groundwater discharges along two 5th order streams, the Housatonic and Farmington Rivers, and 32 1st to 4th order reaches across the Farmington River network. We mapped locations of preferential groundwater discharge exposed along streambanks using handheld thermal infrared cameras paired with high‐resolution topographic and land use land cover datasets, surficial soil characteristic maps, and depth‐to‐bedrock geophysical measurements. The unconfined Housatonic River, MA, USA (12 km) had fewer discharge locations and less lateral extent (41 discharge locations with 38 m of active discharge/km of river) compared to the partially confined Farmington River, CT, USA (26 km; 169 discharge locations with 129 m of active discharge/km of river). Using a moving window analysis, we found along both rivers that discharge was more likely to occur where bank slopes were steeper, floodplain extent was narrower, and degree of confinement was higher. Along the Farmington River, groundwater discharge was more likely to occur where saturated hydraulic conductivity was higher and depth‐to‐bedrock was shallower. Among the 32 stream reaches surveyed (33.2 km of total stream length) within the Farmington River watershed, preferential discharge was observed in all but two stream reaches, varied from 0 to 25% of lateral extent along stream banks (mean = 6%), and was more likely to occur where stream reach slopes were steep, saturated hydraulic conductivity was high, and watershed urbanization was low. Our results show that, though both surface (e.g., topographic, land use land cover) and subsurface (e.g., soil characteristics, bedrock depth) factors control the prevalence of streambank preferential groundwater discharge, the dominant controls vary across valley settings and stream sizes.
Radiogenomics uses machine-learning (ML) to directly connect the morphologic and physiological appearance of tumors on clinical imaging with underlying genomic features. Despite extensive growth in ...the area of radiogenomics across many cancers, and its potential role in advancing clinical decision making, no published studies have directly addressed uncertainty in these model predictions. We developed a radiogenomics ML model to quantify uncertainty using transductive Gaussian Processes (GP) and a unique dataset of 95 image-localized biopsies with spatially matched MRI from 25 untreated Glioblastoma (GBM) patients. The model generated predictions for regional EGFR amplification status (a common and important target in GBM) to resolve the intratumoral genetic heterogeneity across each individual tumor-a key factor for future personalized therapeutic paradigms. The model used probability distributions for each sample prediction to quantify uncertainty, and used transductive learning to reduce the overall uncertainty. We compared predictive accuracy and uncertainty of the transductive learning GP model against a standard GP model using leave-one-patient-out cross validation. Additionally, we used a separate dataset containing 24 image-localized biopsies from 7 high-grade glioma patients to validate the model. Predictive uncertainty informed the likelihood of achieving an accurate sample prediction. When stratifying predictions based on uncertainty, we observed substantially higher performance in the group cohort (75% accuracy, n = 95) and amongst sample predictions with the lowest uncertainty (83% accuracy, n = 72) compared to predictions with higher uncertainty (48% accuracy, n = 23), due largely to data interpolation (rather than extrapolation). On the separate validation set, our model achieved 78% accuracy amongst the sample predictions with lowest uncertainty. We present a novel approach to quantify radiogenomics uncertainty to enhance model performance and clinical interpretability. This should help integrate more reliable radiogenomics models for improved medical decision-making.
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