Protein stability, folding and misfolding in human PGK1 deficiency

Conformational diseases are often caused by mutations, altering protein folding and stability in vivo. We review here our recent work on the effects of mutations on the human phosphoglycerate kinase 1 (hPGK1), with a particular focus on thermodynamics and kinetics of protein folding and misfolding. Expression analyses and in vitro biophysical studies indicate that disease-causing mutations enhance protein aggregation propensity. We found a strong correlation among protein aggregation propensity, thermodynamic stability, cooperativity and dynamics. Comparison of folding and unfolding properties with previous reports in PGKs from other species suggests that hPGK1 is very sensitive to mutations leading to enhance protein aggregation through changes in protein folding cooperativity and the structure of the relevant denaturation transition state for aggregation. Overall, we provide a mechanistic framework for protein misfolding of hPGK1, which is insightful to develop new therapeutic strategies aimed to target native state stability and foldability in hPGK1 deficient patients. © 2013 by the authors; licensee MDPI, Basel, Switzerland

Structural Characterization of Rapid Thermal Oxidized Si\u3csub\u3e1−x−y\u3c/sub\u3eGe\u3csub\u3ex\u3c/sub\u3eC\u3csub\u3ey\u3c/sub\u3e Alloy Films Grown by Rapid Thermal Chemical Vapor Deposition

The structural properties of as-grown and rapid thermal oxidized Si1−x−yGexCy epitaxial layers have been examined using a combination of infrared, x-ray photoelectron, x-ray diffraction, secondary ion mass spectroscopy, and Raman spectroscopy techniques. Carbon incorporation into the Si1−x−yGexCy system can lead to compressive or tensile strain in the film. The structural properties of the oxidized Si1−x−yGexCy film depend on the type of strain (i.e., carbon concentration) of the as-prepared film. For compressive or fully compensated films, the oxidation process drastically reduces the carbon content so that the oxidized films closely resemble to Si1−xGex films. For tensile films, two broad regions, one with carbon content higher and the other lower than that required for full strain compensation, coexist in the oxidized films

Site-to-site interdomain communication may mediate different loss-of-function mechanisms in a cancer-associated NQO1 polymorphism

Disease associated genetic variations often cause intracellular enzyme inactivation, dysregulation and instability. However, allosteric communication of mutational effects to distant functional sites leading to loss-of-function remains poorly understood. We characterize here interdomain site-to-site communication by which a common cancer-associated single nucleotide polymorphism (c.C609T/p.P187S) reduces the activity and stability in vivo of NAD(P)H:quinone oxidoreductase 1 (NQO1). NQO1 is a FAD-dependent, two-domain multifunctional stress protein acting as a Phase II enzyme, activating cancer pro-drugs and stabilizing p53 and p73a oncosuppressors. We show that p.P187S causes structural and dynamic changes communicated to functional sites far from the mutated site, affecting the FAD binding site located at the N-terminal domain (NTD) and accelerating proteasomal degradation through dynamic effects on the C-terminal domain (CTD). Structural protein:protein interaction studies reveal that the cancer-associated polymorphism does not abolish the interaction with p73a, indicating that oncosuppressor destabilization largely mirrors the low intracellular stability of p.P187S. In conclusion, we show how a single disease associated amino acid change may allosterically perturb several functional sites in an oligomeric and multidomain protein. These results have important implications for the understanding of loss-of-function genetic diseases and the identification of novel structural hot spots as targets for pharmacological intervention

Spatially Controlled Generation and Probing of Random Telegraph Noise in Metal Nanocrystal Embedded HfO2Using Defect Nanospectroscopy

Random telegraph noise (RTN) is often considered a nuisance or, more critically, a key reliability challenge for miniaturized semiconductor devices. However, this picture is gradually changing as recent works have shown emerging applications based on the inherent randomness of the RTN signals in state-of-The-Art technologies, including true random number generator and IoT hardware security. Suitable material platforms and device architectures are now actively explored to bring these technologies from an embryonic stage to practical application. A key challenge is to devise material systems, which can be reliably used for the deterministic creation of localized defects to be used for RTN generation. Toward this goal, we have investigated RTN in Au nanocrystal (Au-NC) embedded HfO2stacks at the nanoscale by combining conduction atomic force microscopy defect spectroscopy and a statistical factorial hidden Markov model analysis. With a voltage applied across the stack, there is an enhanced asymmetric electric field surrounding the Au-NC. This in turn leads to the preferential generation of atomic defects in the HfO2near the Au-NC when voltage is applied to the stack to induce dielectric breakdown. Since RTN arises from various electrostatic interactions between closely spaced atomic defects, the Au-NC HfO2material system exhibits an intrinsic ability to generate RTN signals. Our results also highlight that the spatial confinement of multiple defects and the resulting electrostatic interactions between the defects provides a dynamic environment leading to many complex RTN patterns in addition to the presence of the standard two-level RTN signals. The insights obtained at the nanoscale are useful to optimize metal nanocrystal embedded high-κ stacks and circuits for on-demand generation of RTN for emerging random number applications

Fish mitigate trophic depletion in marine cave ecosystems.

Dark marine habitats are often characterized by a food-limited condition. Peculiar dark habitats include marine caves, characterized by the absence of light and limited water flow, which lead to reduced fluxes of organic matter for cave-dwelling organisms. We investigated whether the most abundant and common cave-dwelling fish Apogon imberbis has the potential to play the role of trophic vector in Mediterranean marine caves. We first analysed stomach contents to check whether repletion changes according to a nycthemeral cycle. We then identified the prey items, to see whether they belong to species associated with cave habitats or not. Finally, we assessed whether A. imberbis moves outside marine caves at night to feed, by collecting visual census data on A. imberbis density both inside and outside caves, by day and by night. The stomach repletion of individuals sampled early in the morning was significantly higher than later in the day. Most prey were typical of habitats other than caves. A. imberbis was on average more abundant within caves during the day and outside during the night. Our study supports the hypothesis regarding the crucial trophic role of A. imberbis in connecting Mediterranean marine caves with external habitat

Enhanced vulnerability of human proteins towards disease-associated inactivation through divergent evolution

Human proteins are vulnerable towards disease-associated single amino acid replacements affecting protein stability and function. Interestingly, a few studies have shown that consensus amino acids from mammals or vertebrates can enhance protein stability when incorporated into human proteins. Here, we investigate yet unexplored relationships between the high vulnerability of human proteins towards disease-associated inactivation and recent evolutionary site-specific divergence of stabilizing amino acids. Using phylogenetic, structural and experimental analyses, we show that divergence from the consensus amino acids at several sites during mammalian evolution has caused local protein destabilization in two human proteins linked to disease: cancer-associated NQO1 and alanine: glyoxylate aminotransferase, mutated in primary hyperoxaluria type I. We demonstrate that a single consensus mutation (H80R) acts as a disease suppressor on the most common cancer-associated polymorphism in NQO1 (P187S). The H80R mutation reactivates P187S by enhancing FAD binding affinity through local and dynamic stabilization of its binding site. Furthermore, we show how a second suppressor mutation (E247Q) cooperates with H80R in protecting the P187S polymorphism towards inactivation through long-range allosteric communication within the structural ensemble of the protein. Our results support that recent divergence of consensus amino acids may have occurred with neutral effects on many functional and regulatory traits of wild-type human proteins. However, divergence at certain sites may have increased the propensity of some human proteins towards inactivation due to disease-associated mutations and polymorphisms. Consensus mutations also emerge as a potential strategy to identify structural hot-spots in proteins as targets for pharmacological rescue in loss-of-function genetic diseases.Spanish Ministry of Economy and Competitiveness, MINECO (BIO 2015 66426-R to JMSR, CTQ 2015-64445-R to JLN, ‘Factoría Española de Cristalización’, Consolider-Ingenio 2010 to JAG and SAF2015-69796 to ES), Junta de Andalucia (P11-CTS-07187 to ALP) and FEDER fun

Use of mHealth Technology for Patient-Reported Outcomes in Community-Dwelling Adults with Acquired Brain Injuries: A Scoping Review.

The purpose of our scoping review was to describe the current use of mHealth technology for long-term assessment of patient-reported outcomes in community-dwelling individuals with acquired brain injury (ABI). Following PRISMA guidelines, we conducted a scoping review of literature meeting these criteria: (1) civilians or military veterans, all ages; (2) self-reported or caregiver-reported outcomes assessed via mobile device in the community (not exclusively clinic/hospital); (3) published in English; (4) published in 2015-2019. We searched Ovid MEDLINE(R) \u3c 1946 to 16 August 2019, MEDLINE InProcess, EPub, Embase, and PsycINFO databases for articles. Thirteen manuscripts representing 12 distinct studies were organized by type of ABI [traumatic brain injury (TBI) and stroke] to extract outcomes, mHealth technology used, design, and inclusion of ecological momentary assessment (EMA). Outcomes included post-concussive, depressive, and affective symptoms, fatigue, daily activities, stroke risk factors, and cognitive exertion. Overall, collecting patient-reported outcomes via mHealth was feasible and acceptable in the chronic ABI population. Studies consistently showed advantage for using EMA despite variability in EMA timing/schedules. To ensure best clinical measurement, research on post-ABI outcomes should consider EMA designs (versus single time-point assessments) that provide the best timing schedules for their respective aims and outcomes and that leverage mHealth for data collection

Posidonia oceanica restoration, a relevant strategy after boat anchoring degradation?

Trabajo presentado al 7th Mediterranean Symposium on Marine Vegetation, celebrado en Génova (Italia), los días 19 y 20 de septiembre de 2022.The anchoring of large pleasure boats constitutes one of the main threats in shallow marine habitats and particularly for seagrass beds. In the Mediterranean, this activity has seen constant development during the last decades, causing major physical disturbances in Posidonia oceanica meadows and associated ecosystem services, notably in terms of climate change mitigation (i.e. carbon fixation and sequestration capacities). In this context, the aims of the present study are to estimate the impact of these anchoring activities on P. oceanica meadows in a particularly highlyfrequented area (Sant’Amanza gulf, SE Corsica Island) and to set up a strategy to restore this major carbon sink. Since the last decade, time-series of marine habitat maps revealed an important regression of P. oceanica meadows, with a loss of 72.9 ha, corresponding to 11% of the meadow surface and 9% decline in the total carbon fixation and sequestration performed each year. Moreover, in the most impacted part of the bay (Balistra bay), a loss of 16.6 ha (28%) has been recorded between 2011 and 2022. Following recent enforcement of anchoring regulation, prohibiting the anchoring of large units (greater than 24 m), and the lockdown linked to the COVID-19 pandemic, anchorages in the seagrass reduced by 92%, between 2018 and 2022. Natural recovery was observed at the edge of the meadow (plagiotropic rhizomes) but this growth is slow and the areas to recover are large. As a result, a transplant experiment, from cuttings harvested from the adjacent meadows, was initiated in the spring 2021. Four restoration techniques are being tested in the some pilot sites and the development of these transplants will be monitored and compared to the natural recovery.This research has been co-financed by INTERREG program (GIREPAM), Collectivity of Corsica Corse (PADDUC-CHANGE), SETEC Foundation, Environment Office of Corsica and the French Office of Biodiversity (RenforC program)

The regulatory subunit of PKA-I remains partially structured and undergoes β-aggregation upon thermal denaturation

Background: The regulatory subunit (R) of cAMP-dependent protein kinase (PKA) is a modular flexible protein that responds with large conformational changes to the binding of the effector cAMP. Considering its highly dynamic nature, the protein is rather stable. We studied the thermal denaturation of full-length RIα and a truncated RIα(92-381) that contains the tandem cyclic nucleotide binding (CNB) domains A and B. Methodology/Principal Findings: As revealed by circular dichroism (CD) and differential scanning calorimetry, both RIα proteins contain significant residual structure in the heat-denatured state. As evidenced by CD, the predominantly α-helical spectrum at 25°C with double negative peaks at 209 and 222 nm changes to a spectrum with a single negative peak at 212-216 nm, characteristic of β-structure. A similar α→β transition occurs at higher temperature in the presence of cAMP. Thioflavin T fluorescence and atomic force microscopy studies support the notion that the structural transition is associated with cross-β-intermolecular aggregation and formation of non-fibrillar oligomers. Conclusions/Significance: Thermal denaturation of RIα leads to partial loss of native packing with exposure of aggregation-prone motifs, such as the B' helices in the phosphate-binding cassettes of both CNB domains. The topology of the β-sandwiches in these domains favors inter-molecular β-aggregation, which is suppressed in the ligand-bound states of RIα under physiological conditions. Moreover, our results reveal that the CNB domains persist as structural cores through heat-denaturation. © 2011 Dao et al