Crowding-induced protein destabilization in the absence of soft attractions

It is generally assumed that volume exclusion by macromolecular crowders universally stabilizes the native states of proteins and destabilization suggests soft attractions between crowders and protein. Here we show that proteins can be destabilized even by crowders that are purely repulsive. With a coarse-grained sequence-based model, we study the folding thermodynamics of two sequences with different native folds, a helical hairpin and a β-barrel, in a range of crowder volume fractions, ${\phi }_{\text{c}}$. We find that the native state, N, remains structurally unchanged under crowded conditions, while the size of the unfolded state, U, decreases monotonically with ${\phi }_{\text{c}}$. Hence, for all ${\phi }_{\text{c}}>0$, U is entropically disfavored relative to N. This entropy-centric view holds for the helical hairpin protein, which is stabilized under all crowded conditions as quantified by changes in either the folding midpoint temperature, $T_{\text{m}}$, or the free energy of folding. We find, however, that the β-barrel protein is destabilized under low-T, low-${\phi }_{\text{c}}$ conditions. This destabilization can be understood from two characteristics of its folding: 1) a relatively compact U at $T<T_{\text{m}}$, such that U is only weakly disfavored entropically by the crowders; and 2) a transient, compact, and relatively low-energy nonnative state that has a maximum population of only a few percent at ${\phi }_{\text{c}}=0$, but increasing monotonically with ${\phi }_{\text{c}}$. Overall, protein destabilization driven by hard-core effects appears possible when a compaction of U leads to even a modest population of compact nonnative states that are energetically competitive with N.     

Significant compaction of H4 histone tail upon charge neutralization by acetylation and its mimics,possible effects on chromatin structure

The intrinsically disordered, positively charged H4 histone tail is important for chromatin structure and function. We have explored conformational ensembles of human H4 tail in solution, with varying levels of charge neutralization via acetylation or amino-acid substitutions such as $K\to Q$. We have employed an explicit water model shown recently to be well suited for simulations of intrinsically disordered proteins.Upon progressive neutralization of the H4, its radius of gyration decreases linearly with the tail charge q, the trend is explained using a simple polymer model. While the wild type state ($q=+8$) is essentially a random coil, hyper-acetylated H4 ($q=+3$) is virtually as compact and stable as a globular protein of the same number of amino-acids.Conformational ensembles of acetylated H4 match the corresponding $K\to X$ substitutions only approximately: based on the ensemble similarity, we propose $K\to M$ as a possible alternative to the commonly used $K\to Q$.Possible effects of the H4 tail compaction on chromatin structure are discussed within a qualitative model in which the chromatin is highly heterogeneous, easily inter-converting between various structural forms. We predict that upon progressive charge neutralization of the H4 tail, the least compact sub-states of chromatin de-condense first, followed by de-condensation of more compact structures, e.g. those that harbor a high fraction of stacked di-nucleosomes. The predicted hierarchy of DNA accessibility increase upon progressive acetylation of H4 might be utilized by the cell for selective DNA accessibility control.     

On the design principles of metabolic flux sensing

Metabolism is precisely coordinated, with the goal of balancing fluxes to maintain robust growth. However, coordinating fluxes requires information about rates, which can only be inferred through concentrations. While flux-sensitive metabolites have been reported, the design principles underlying such sensing have not been clearly elucidated. Here we use kinetic modeling to show that substrate concentrations of thermodynamically constrained reactions reflect upstream flux and therefore carry information about rates. Then we use untargeted multi-omic data from Escherichia coli and Saccharomyces cerevisiae to show that the concentrations of some metabolites in central carbon metabolism reflect fluxes as a result of thermodynamic constraints. We then establish, using 37 real concentration-flux relationships across both organisms, that in vivo $\text{Δ}{G}^{°}\ge ?4$ kJ/mol is the threshold above which substrates are likely to be sensitive to upstream flux(es).     

Implications of critical node-dependent unidirectional cross-talk of Plasmodium SUMO pathway proteins

The endoparasitic pathogen, Plasmodium falciparum (Pf), modulates protein-protein interactions to employ post-translational modifications like SUMOylation to establish successful infections. The interaction between E1 and E2 (Ubc9) enzymes governs species specificity in the Plasmodium SUMOylation pathway. Here, we demonstrate that a unidirectional cross-species interaction exists between Pf-SUMO and human E2, whereas Hs-SUMO1 failed to interact with Pf-E2. Biochemical and biophysical analyses revealed that surface-accessible aspartates of Pf-SUMO determine the efficacy and specificity of SUMO-Ubc9 interactions. Furthermore, we demonstrate that critical residues of the Pf-Ubc9 N terminus are responsible for diminished Hs-SUMO1 and Pf-Ubc9 interaction. Mutating these residues to corresponding Hs-Ubc9 residues restores electrostatic, $\pi ?\pi$, and hydrophobic interactions and allows efficient cross-species interactions. We suggest that, in comparison with human counterparts, Plasmodium SUMO and Ubc9 proteins have acquired critical changes on their surfaces as nodes, which Plasmodium can use to exploit the host SUMOylation machinery.     

Spontaneous local membrane curvature induced by transmembrane proteins

The (local) curvature of cellular membranes acts as a driving force for the targeting of membrane-associated proteins to specific membrane domains, as well as a sorting mechanism for transmembrane proteins, e.g., by accumulation in regions of matching spontaneous curvature. The latter measure was previously experimentally employed to study the curvature induced by the potassium channel KvAP and by aquaporin AQP0. However, the direction of the reported spontaneous curvature levels as well as the molecular driving forces governing the membrane curvature induced by these integral transmembrane proteins could not be addressed experimentally.Here, using both coarse-grained and atomistic molecular dynamics (MD) simulations, we report induced spontaneous curvature values for the homologous potassium channel Kv 1.2/2.1 Chimera (KvChim) and AQP0 embedded in unrestrained lipid bicelles that are in very good agreement with experiment. Importantly, the direction of curvature could be directly assessed from our simulations: KvChim induces a strong positive membrane curvature ($\approx 0.036$ nm^?1) whereas AQP0 causes a comparably small negative curvature ($\approx ?0.019$ nm^?1).Analyses of protein-lipid interactions within the bicelle revealed that the potassium channel shapes the surrounding membrane via structural determinants. Differences in shape of the protein-lipid interface of the voltage-gating domains between the extracellular and cytosolic membrane leaflets induce membrane stress and thereby promote a protein-proximal membrane curvature. In contrast, the water pore AQP0 displayed a high structural stability and an only faint effect on the surrounding membrane environment that is connected to its wedge-like shape.     

A New Compartmental Epidemiological Model for COVID-19 with a Case Study of Portugal

We propose a compartmental mathematical model for the spread of the COVID-19 disease, showing its usefulness with respect to the pandemic in Portugal, from the first recorded case in the country till the end of the three states of emergency. New results include the compartmental model, described by a system of seven ordinary differential equations; proof of positivity and boundedness of solutions; investigation of equilibrium points and their stability analysis; computation of the basic reproduction number; and numerical simulations with official real data from the Portuguese health authorities. Besides completely new, the proposed model allows to describe quite well the spread of COVID-19 in Portugal, fitting simultaneously not only the number of active infected individuals but also the number of hospitalized individuals, respectively with a $L^2$ error of $9.2152e-04$ and $1.6136e-04$ with respect to the initial population. Such results are very important, from a practical point of view, and far from trivial from a mathematical perspective. Moreover, the obtained value for the basic reproduction number is in agreement with the one given by the Portuguese authorities at the end of the three emergency states.     

Penicillium rubens germination on desiccated and nutrient-depleted conditions depends on the water activity during sporogenesis

Fungal growth often appears in a surrounding where water and nutrients are scarce. The impact of this environment during sporogenesis on subsequent growth is often neglected.This study investigates the effect of water availability during sporogenesis on subsequent early growth. Therefore, a carbon-depleted substrate was constructed. Humidity is then the only parameter of interest. The water conditions during sporogenesis, and during subsequent growth, were varied. This is a stressing environment: no carbon source is present, and water provided solely via the vapour.The lag time, $t_l$, and initial growth rate, ${\mu }_{fp}$, of the germ tubes were monitored.The effect of $a_w$ history on germination and initial growth depends on the $RH$ of the environment. Only at low $RH$ do spores produced at low $a_w$ have a smaller $t_l$ and higher ${\mu }_{fp}$ compared to those grown at high $a_w$. This result was remarkably pronounced when the substrate was also made hydrophobic: growth only occurred when spores were developed at low $a_w$ and placed in high $RH$.Spores grown on lowered $a_w$ attract more water. It is hypothesized that this attraction affects subsequent growth behaviour, and is the reason why growth on hydrophobic glass only prevails in the condition of high $RH$ and lowered $a_w$ history.We demonstrate the influence of cultivation conditions on germination, which becomes more pronounced in a more desiccated environment.