Protein thermostability above 100 degreesC: a key role for ionic interactions

The discovery of hyperthermophilic microorganisms and the analysis of hyperthermostable enzymes has established the fact that multisubunit enzymes can survive for prolonged periods at temperatures above 100 degreesC. We have carried out homology-based modeling and direct structure comparison on the hexameric glutamate dehydrogenases from the hyperthermophiles Pyrococcus furiosus and Thermococcus litoralis whose optimal growth temperatures are 100 degreesC and 88 degreesC, respectively, to determine key stabilizing features. These enzymes, which are 87% homologous, differ 16-fold in thermal stability at 104 degreesC. We observed that an intersubunit ion-pair network was substantially reduced in the less stable enzyme from T. litoralis, and two residues were then altered to restore these interactions. The single mutations both had adverse effects on the thermostability of the protein. However, with both mutations in place, we observed a fourfold improvement of stability at 104 degreesC over the wild-type enzyme. The catalytic properties of the enzymes were unaffected by the mutations. These results suggest that extensive ion-pair networks may provide a general strategy for manipulating enzyme thermostability of multisubunit enzymes. However, this study emphasizes the importance of the exact local environment of a residue in determining its effects on stability.     

The Homothorax homeoprotein activates the nuclear localization of another homeoprotein, extradenticle, and suppresses eye development in Drosophila

The Extradenticle (Exd) protein in Drosophila acts as a cofactor to homeotic proteins. Its nuclear localization is regulated. We report the cloning of the Drosophila homothorax (hth) gene, a homolog of the mouse Meis1 proto-oncogene that has a homeobox related to that of exd. Comparison with Meis1 finds two regions of high homology: a novel MH domain and the homeodomain. In imaginal discs, hth expression coincides with nuclear Exd. hth and exd also have virtually identical, mutant clonal phenotypes in adults. These results suggest that hth and exd function in the same pathway. We show that hth acts upstream of exd and is required and sufficient for Exd protein nuclear localization. We also show that hth and exd are both negative regulators of eye development; their mutant clones caused ectopic eye formation. Targeted expression of hth, but not of exd, in the eye disc abolished eye development completely. We suggest that hth acts with exd to delimit the eye field and prevent inappropriate eye development.     

Identification of Entry Inhibitors against Delta and Omicron Variants of SARS-CoV-2

Entry inhibitors against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are urgently needed to control the outbreak of coronavirus disease 2019 (COVID-19). This study developed a robust and straightforward assay that detected the molecular interaction between the receptor-binding domain (RBD) of viral spike protein and the angiotensin-converting enzyme 2 (ACE2) receptor in just 10 min. A drug library of 1068 approved compounds was used to screen for SARS-CoV2 entry inhibition, and 9 active drugs were identified as specific pseudovirus entry inhibitors. A plaque reduction neutralization test using authentic SARS-CoV-2 virus in Vero E6 cells confirmed that 2 of these drugs (Etravirine and Dolutegravir) significantly inhibited the infection of SARS-CoV-2. With molecular docking, we showed that both Etravirine and Dolutegravir are preferentially bound to primary ACE2-interacting residues on the RBD domain, implying that these two drug blocks may prohibit the viral attachment of SARS-CoV-2. We compared the neutralizing activities of these entry inhibitors against different pseudoviruses carrying spike proteins from alpha, beta, gamma, and delta variants. Both Etravirine and Dolutegravir showed similar neutralizing activities against different variants, with EC50 values between 4.5 to 5.8 nM for Etravirine and 10.2 to 22.9 nM for Dolutegravir. These data implied that Etravirine and Dolutegravir may serve as general spike inhibitors against dominant viral variants of SARS-CoV-2.     

Effect of silanes on the morphology,hydrophobicity, and dynamical mechanical properties of polyimide/silica hybrid membranes

Polyimide (PI)/silica hybrid membranes with high contact angles were prepared through the in situ sol–gel process. The precursor, poly(amic acid) with controlled block chain length, was synthesized using 4,4′‐diaminodiphenyl ether (ODA), 3,3′,4,4′‐benzophenone‐tetracarboxylic dianhydride (BTDA) and 3‐aminopropyl‐trimethoxysilane (APrTMOS) or 3‐aminopropyldimethylethoxysilane (APDiMOS). And then, phenyltrimethoxysilane (PTS) or tetramethoxysilane (TMOS) or methyltrimethoxysilane (MTrMOS) was respectively, added to the above polyamic acid and mixed thoroughly. Following curing reaction, the PI/silica hybrid membranes with different cross‐linkages, silica content, and hydrophobic properties were prepared. The effect on the formation of PI imide ring during imidization reaction is increased as the increase of silanes content and characterized by frequency shiftment and absorbance ratio of Fourier transform infrared (FTIR) measurements. All the hybrid membranes show high transparency though with high silica contents. The storage modulus, tan δ, and damping intensity by DMA measurements are all correlated with silane content or block chain length. And all these membranes with silane content possess high contact angle as compared to pure PI without any silanes added and the contact angles increase with increasing the silane content. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Torrefied biomass‐polypropylene composites

Torrefied almond shells and wood chips were incorporated into polypropylene as fillers to produce torrefied biomass‐polymer composites. The composites were prepared by extrusion and injection molding. Response surface methodology was used to examine the effects of filler concentration, filler size, and lignin factor (relative lignin to cellulose concentration) on the material properties of the composites. The heat distortion temperatures, thermal properties, and tensile properties of the composites were characterized by thermomechanical analysis, differential scanning calorimetry, and tensile tests, respectively. The torrefied biomass composites had heat distortion temperatures of 8–24°C higher than that of neat polypropylene. This was due to the torrefied biomass restricting mobility of polypropylene chains, leading to higher temperatures for deformation. The incorporation of torrefied biomass generally resulted in an increase in glass transition temperature, but did not affect melting temperature. Also, the composites had lower tensile strength and elongation at break values than those of neat polypropylene, indicating weak adhesion between torrefied biomass and polypropylene. However, scanning electron microscopy results did indicate some adhesion between torrefied biomass and polypropylene. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41582.     

Bioactive Chemical Constituents from the Brown Alga Homoeostrichus formosana

A new chromene derivative, 2-(4'',8''-dimethylnona-3''E,7''-dienyl)-8-hydroxy-2,6-dimethyl-2H-chromene (1) together with four known natural products, methylfarnesylquinone (2), isololiolide (3), pheophytin a (4), and β-carotene (5) were isolated from the brown alga Homoeostrichus formosana. The structure of 1 was determined by extensive 1D and 2D spectroscopic analyses. Acetylation of 1 yielded the monoacetylated derivative 2-(4'',8''-dimethylnona-3''E,7''-dienyl)-8-acetyl-2,6-dimethyl-2H-chromene (6). Compounds 1–6 exhibited various levels of cytotoxic, antibacterial, and anti-inflammatory activities. Compound 2 was found to display potent in vitro anti-inflammatory activity by inhibiting the generation of superoxide anion (IC₅₀ 0.22 ± 0.03 μg/mL) and elastase release (IC₅₀ 0.48 ± 0.11 μg/mL) in FMLP/CB-induced human neutrophils.     

Effect of electrochemical surface area on electroless copper deposition: Pd nanocubes as new activators

In this study, the electrochemical surface area (ESA) and specific activity are used to obtain a fair comparison of various activators used for electroless copper deposition (ECD). Different sizes (i.e., 27, 48, and 63 nm) of Pd nanocubes (NCs) enclosed by {1 0 0} facets were successfully synthesized for use as new activators. The results of an analysis based on mixed potential theory indicated that the specific activities in terms of deposition current densities for the 27, 48, and 63 nm NCs and 9 nm nanoparticles (NPs) were 0.31, 1.06, 0.93, and 0.13 mA cm^?2_ESA, respectively. From the results of electrochemical quartz crystal microbalance measurements, the specific activities calculated from the mean deposition rates of the 27, 48, and 63 nm NCs and 9 nm NPs were 29.62, 68.72, 54.77, and 9.72 μg cm^?2 s^?1, respectively. A specific activity order of 48 nm NC > 63 nm NC > 27 nm NC > 9 nm NP was obtained. The 48 nm Pd NCs exhibited the maximum activity towards the activation of the ECD bath.     

Low-Temperature Sintering and Microwave Dielectric Properties of Anorthite-Based Glass-Ceramics

TiO₂ nucleated anorthite-based glass-ceramics were fabricated from glass powders. After sintering and crystallization heat treatment, various physical properties, including apparent bulk density and water absorption, were examined to evaluate the sintering behavior of anorthite-based glass-ceramic. Results showed that the complete-densification temperature for specimens was as low as 900°C. Sufficient crystallization was achieved by subsequently raising the firing temperature to 950°C, and the dielectric quality factor was promoted to the maximum value. Contents of nucleating agent (TiO₂) played an important role in the dielectric constants. The crystallinity was controlled by raising the firing temperature at a constant heating rate. The degree of crystallization affected the dielectric properties of sintered glass-ceramics. At the resonant frequency of 10 GHz, anorthite glass-ceramics with 5 wt% TiO₂ possessed the lowest permittivity of 8 and exhibited appropriate dielectric properties as compared with those with B₂O₃ and 10 wt% TiO₂.     

Structural and functional analogy between pneumolysin and proaerolysin

Pneumolysin and proaerolysin are bacterial toxins that form pores in host cells by oligomerization. We propose that they may have similar structures despite a poor sequence identity. The crystal structure of proaerolysin reveals a protein composed of four domains, arranged in the shape of an elongated comma. Electron microscopy of the pneumolysin monomer shows a similar arrangement of domains. The sequence of pneumolysin recognizes the template of proaerolysin from a library of protein folds. A three-dimensional model of pneumolysin has been constructed by the comparative approach using the structure of proaerolysin. This model, together with results on the activity of site- specific mutants and the positions of antigenic sites, has been used to propose functional roles of individual domains.