ABSTRACT
Non-biodegradable polyolefin based plastic mulch residues in agricultural fields after the end of a crop cycle have raised several concerns as an environmental pollutant in recent years. This study explores the potential of Poly (lactic acid) (PLA) and Poly (butylene adipate-co-terephthalate) (PBAT) based compostable films reactively blended with compatibilizers and chain extenders as a promising solution to environmental challenges associated with traditional plastic mulch films. Epoxidized soybean oil (ESO) and Epoxy-functionalized styrene acrylic copolymer (ESA) have been used as reactive compatibilizers and chain extenders respectively. In-depth analysis of the mechanical, thermal, and barrier properties of the developed films, revealed that the PLA/PBAT blend films at 75:25 weight ratio in the presence of 5 phr ESO and 0.5 phr ESA exhibit improved performance characteristics for application as mulch films. Furthermore, the films were subjected to 360-h UV exposure to gauge their stability under prolonged exposure, specifically investigating changes in the carbonyl index. Additionally, a rigorous real-time field trial of the mulch films spanning eight months with various crops was carried out to understand their performance in practical agricultural settings. The study also involved the identification of microorganisms responsible for the degradation of the developed mulch films employing 16S rRNA sequencing.
Subject(s)
Agriculture , Biodegradation, Environmental , Polyesters , Soil Microbiology , Polyesters/chemistry , Agriculture/methods , Microbiota , Soil/chemistry , Soybean OilABSTRACT
OBJECTIVE: Designing a new clinical trial entails many decisions, such as defining a cohort and setting the study objectives to name a few, and therefore can benefit from recommendations based on exhaustive mining of past clinical trial records. This study proposes an approach based on knowledge graph embeddings and semantics-driven inductive inference for generating such recommendations. METHOD: The proposed recommendation methodology is based on neural embeddings trained on first-of-its-kind knowledge graph constructed from clinical trials data. The methodology includes design of a knowledge graph for clinical trial data, evaluation of various knowledge graph embedding techniques for it, application of a novel inductive inference method using these embeddings, and generation of recommendations for clinical trial design. The study uses freely available data from clinicaltrials.gov and related sources. RESULTS: The proposed approach for recommendations obtained relevance scores ranging from 70% to 83%. These scores were determined by evaluating the text similarity of recommended elements to actual elements used in clinical trials that are in progress. Furthermore, the most pertinent recommendations were consistently located towards the top of the list, indicating the effectiveness of our method. CONCLUSION: Our study suggests that inductive inference using node semantics is a viable approach for generating recommendations using graphs neural embeddings, and that there is a potential for improvement in training graph embeddings using node semantics.
Subject(s)
Clinical Trials as Topic , Semantics , Humans , Data Mining/methods , Algorithms , Neural Networks, Computer , Research DesignABSTRACT
N-linked glycosylation is an essential and highly conserved co- and post-translational protein modification in all domains of life. In humans, genetic defects in N-linked glycosylation pathways result in metabolic diseases collectively called Congenital Disorders of Glycosylation. In this modification reaction, a mannose rich oligosaccharide is transferred from a lipid-linked donor substrate to a specific asparagine side-chain within the -N-X-T/S- sequence (where X ≠ Proline) of the nascent protein. Oligosaccharyltransferase (OST), a multi-subunit membrane embedded enzyme catalyzes this glycosylation reaction in eukaryotes. In yeast, Ost4 is the smallest of nine subunits and bridges the interaction of the catalytic subunit, Stt3, with Ost3 (or its homolog, Ost6). Mutations of any C-terminal hydrophobic residues in Ost4 to a charged residue destabilizes the enzyme and negatively impacts its function. Specifically, the V23D mutation results in a temperature-sensitive phenotype in yeast. Here, we report the reconstitution of both purified recombinant Ost4 and Ost4V23D each in a POPC/POPE lipid bilayer and their resonance assignments using heteronuclear 2D and 3D solid-state NMR with magic-angle spinning. The chemical shifts of Ost4 changed significantly upon the V23D mutation, suggesting a dramatic change in its chemical environment.
Subject(s)
Hexosyltransferases , Liposomes , Membrane Proteins , Nuclear Magnetic Resonance, Biomolecular , Hexosyltransferases/genetics , Hexosyltransferases/chemistry , Hexosyltransferases/metabolism , Nuclear Magnetic Resonance, Biomolecular/methods , Membrane Proteins/chemistry , Membrane Proteins/genetics , Membrane Proteins/metabolism , Liposomes/chemistry , Saccharomyces cerevisiae Proteins/chemistry , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Mutation , Glycosylation , Protein Subunits/chemistry , Protein Subunits/geneticsABSTRACT
Ostrinia nubilalis, also known as European Corn Borer (ECB), is a serious pest in Europe and North America, as well as in Central Asia and Northern Africa. It damages a variety of agricultural crops such as corn, oats, buckwheat, millet, and soybeans. causing annually at least one billion dollars in loss. The Ostrinia nubilalis pheromone-binding protein3 (OnubPBP3), preferentially expressed in the male moth antenna, has been implicated in the detection of the female-secreted pheromone blend during the mating process. Understanding the structure of and function of OnubPBP3, including the mechanism of pheromone binding and its release at the dendritic olfactory neuron (ORN), is essential if integrated pest management through sensory inhibition is to be achieved. We report here the backbone and side-chain resonance assignments of OnubPBP3 at pH 6.5 using various triple resonance NMR experiments on a 13C, 15N-labeled protein sample. The secondary structure of OnubPBP3 consists of six α-helices and an unstructured C-terminus based on backbone chemical shifts.
Subject(s)
Moths , Sex Attractants , Animals , Male , Female , Sex Attractants/chemistry , Sex Attractants/pharmacology , Sex Attractants/physiology , Nuclear Magnetic Resonance, Biomolecular , Moths/metabolism , Pheromones , AgricultureABSTRACT
The growing popularity of additive manufacturing in the science, industry is associated with high-quality products for futuristic applications. This study presents an in-depth characterization and analysis of the effect of poly (ethylene glycol) (PEG) having molecular weight 6000 g/mol used with various concentrations (1%,3%,5%) to modify the 3D printed Polylactide (PLA) part. The influence of PEG on the morphology, structure, thermal, wettability and mechanical properties of the 3D-printed PLA/PEG part was investigated. Herein, the mechanical property of injection moulding, 3D printed specimens, and finite element analysis (FEA) simulation results were also compared. The structure and properties of PLA/PEG blends were different from those of virgin PLA. By DSC analysis, it was found that the glass transition temperature (Tg) and cold crystallization temperature decreased in the case of the PLA/PEG blend. From TGA it was observed that PLA/PEG blend was thermally stable. It was shown that with the addition of PEG into PLA the tensile strength and young's modulus decrease, whereas elongation percentage and impact strength increase predominantly. The contact angle results indicate that the addition of PEG lowers the contact angle value of the PLA/PEG blend (from 69.32 ± 1.4° to 45.67 ± 1.2°) and increases surface wettability. With 5% PEG loading, PLA/PEG blend showed optimum structural and mechanical properties together with simple processibility.
Subject(s)
Polyesters , Polyethylene Glycols , Polyethylene Glycols/chemistry , Polyesters/chemistry , Temperature , Printing, Three-DimensionalABSTRACT
Ostrinia furnacalis is an invasive lepidopteran agricultural pest that relies on olfaction for mating and reproduction. Male moths have an extremely sensitive olfactory system that can detect the sex pheromones emitted by females over a great distance. Pheromone-binding proteins present in the male moth antenna play a key role in the pheromone uptake, transport, and release at the dendritic membrane of the olfactory neuron. Here, we report the first high-resolution NMR structure of a pheromone-binding protein from an Ostrinia species at pH 6.5. The core of the Ostrinia furnacalis PBP2 (OfurPBP2) consists of six helices, α1a (2-14), α1b (16-22), α2 (27-37), α3 (46-60), α4 (70-80), α5 (84-100), and α6 (107-124) surrounding a large hydrophobic pocket. The structure is stabilized by three disulfide bridges, 19-54, 50-108, and 97-117. In contrast to the unstructured C-terminus of other lepidopteran PBPs, the C-terminus of OfurPBP2 folds into an α-helix (α7) at pH 6.5. The protein has nanomolar affinity towards both pheromone isomers. Molecular docking of both pheromones, E-12 and Z-12-tetradecenyl acetate, to OfurPBP2 revealed that the residues Met5, Lys6, Met8, Thr9, Phe12, Phe36, Trp37, Phe76, Ser115, Phe118, Lys119, Ile122, His123, and Ala128 interact with both isomers, while Thr9 formed a hydrogen bond with the acetate head group. NMR structure and thermal unfolding studies with CD suggest that ligand release at pH 4.5 is likely due to the partial unfolding of the protein.
Subject(s)
Moths , Sex Attractants , Animals , Carrier Proteins/chemistry , Disulfides/metabolism , Female , Insect Proteins/chemistry , Ligands , Male , Molecular Docking Simulation , Moths/chemistry , Moths/metabolism , Pheromones/chemistry , Pheromones/metabolism , Sex Attractants/metabolismABSTRACT
As the highest percentage of global mortality is caused by several cardiovascular diseases (CVD), maintenance and monitoring of a healthy cardiovascular condition have become the primary concern of each and every individual. Simultaneously, recent progress and advances in wearable pressure sensor technology have provided many pathways to monitor and detect underlying cardiovascular illness in terms of irregularities in heart rate, blood pressure, and blood oxygen saturation. These pressure sensors can be comfortably attached onto human skin or can be implanted on the surface of vascular grafts for uninterrupted monitoring of arterial blood pressure. While the traditional monitoring systems are time-consuming, expensive, and not user-friendly, flexible sensor technology has emerged as a promising and dynamic practice to collect important health information at a comparatively low cost in a reliable and user-friendly way. This Review explores the importance and necessity of cardiovascular health monitoring while emphasizing the role of flexible pressure sensors in monitoring patients' health conditions to avoid adverse effects. A comprehensive discussion on the current research progress along with the real-time impact and accessibility of pressure sensors developed for cardiovascular health monitoring applications has been provided.
Subject(s)
Wearable Electronic Devices , Humans , Monitoring, PhysiologicABSTRACT
Calcium (Ca2+) is well known as a second messenger in eukaryotes, where Ca2+ signaling controls life-sustaining cellular processes. Although bacteria produce the components required for Ca2+ signaling, little is known about the mechanisms of bacterial Ca2+ signaling. Previously, we have identified a putative Ca2+-binding protein EfhP (PA4107) with two canonical EF-hand motifs and reported that EfhP mediates Ca2+ regulation of virulence factors production and infectivity in Pseudomonas aeruginosa, a human pathogen causing life-threatening infections. Here, we show that EfhP selectively binds Ca2+ with 13.7 µM affinity, and that mutations at the +X and -Z positions within each or both EF-hand motifs abolished Ca2+ binding. We also show that the hydrophobicity of EfhP increased in a Ca2+-dependent manner, however no such response was detected in the mutated proteins. 15 N-NMR showed Ca2+-dependent chemical shifts in EfhP confirming Ca2+-binding triggered structural rearrangements in the protein. Deletion of efhP impaired P. aeruginosa survival in macrophages and virulence in vivo. Disabling EfhP Ca2+ binding abolished Ca2+ induction of pyocyanin production in vitro. These data confirm that EfhP selectively binds Ca2+, which triggers its structural changes required for the Ca2+ regulation of P. aeruginosa virulence, thus establishing the role of EfhP as a Ca2+ sensor.
Subject(s)
EF Hand Motifs , Pseudomonas aeruginosa , Calcium/metabolism , Humans , Pseudomonas aeruginosa/physiology , Pyocyanine/metabolism , Virulence , Virulence Factors/genetics , Virulence Factors/metabolismABSTRACT
Improper handling of plastic waste and related chemical pollution has garnered much attention in recent years owing to the associated detrimental impacts on human health and the environment. This article reports an overview of the main interlinkages between persistent organic pollutants (POPs) and plastic in the waste management system of India. Both plastics and POPs share certain common traits such as persistence, resistance to biological degradation, and the ability to get transported over long distances. Throughout the processes of production, consumption, and disposal, plastics interact with and accumulate POPs through several mechanisms and end up co-existing in the environment. Plastic waste can undergo long-range transport through rivers and the oceans, break down into microplastics and get transported through the air, or remain locked in waste dump yards and landfills. Over time, environmental processes lead to the leaching and release of accumulated POPs from these plastic wastes. Plastic recycling in the Indian informal sector including smelting, scrubbing, and shredding of plastic waste, is also a potential major POPs source that demands further investigation. The presence of POPs in plastic waste and their fate in the plastic recycling process have not yet been elucidated. By enhancing our understanding of these processes, this paper may aid policy decisions to combat the release of POPs from different waste types and processes in India.
Subject(s)
Environmental Pollutants , Waste Management , Humans , Plastics , Persistent Organic Pollutants , Environmental Pollution , RecyclingABSTRACT
Introduction: Fibroblast growth factor 23 (FGF-23) is a phosphate metabolism regulator in patients with chronic kidney disease (CKD). The present study is aimed to examine the FGF-23 level in pre-dialysis patients with CKD and its correlation with carotid artery calcification (CAAC). Methods: This cross-sectional study included patients with CKD and controls. The patients were compared with controls having similar distribution of age and sex to determine serum FGF-23 level in Indian healthy adult population. Detailed medical history, physical examination, and investigations were done for each patient. Atherosclerotic risk factors, cardiovascular comorbidities, and drug history were recorded. Carotid calcification was observed using carotid ultrasound. Results: In total, 62 patients with a mean age of 50.0 years were enrolled. Majority of the patients had hypertension (66.1%), followed by diabetes (27.4%) and dyslipidemia (3.2%). Mean serum corrected calcium levels were significantly higher in patients with CAAC compared to the patients without CAAC (9.21 ± 1.34 vs. 8.53 ± 0.93 mg/dL; P = 0.014). The FGF-23 levels were significantly higher in patients with CAAC compared to those without CAAC (396.0 vs. 254.0 pg/mL; P = 0.008). CAAC was found to be present in both early and late stages of CKD. Multivariate analysis showed that log FGF-23 and serum corrected calcium remained as independent determinants of CAAC. The prevalence of CAAC increased with the ascending quartiles of FGF23. Conclusion: In conclusion, FGF-23 was found to be independently associated with CAAC in CKD.
ABSTRACT
Ostrinia nubilalis, a lepidopteran moth, also known as the European corn borer, has a major impact on the production of economically important crops in the United States and Europe. The female moth invites the male moth for mating through the release of pheromones, a volatile chemical signal. Pheromone binding proteins (PBPs) present in the male moth antennae are believed to pick up the pheromones, transport them across the aqueous sensillum lymph, and deliver them to the olfactory receptor neurons. Here we report for the first time the cloning, expression, refolding, purification, and structural characterization of Ostrinia nubilalis PBP3 (OnubPBP3). The recombinant protein showed nanomolar affinity to each isomer of the Ostrinia pheromones, E- and Z-11-tetradecenyl acetate. In a pH titration study by nuclear magnetic resonance, the protein exhibited an acid-induced unfolding at pH below 5.5. The molecular dynamics simulation study demonstrated ligand-induced conformational changes in the protein with both E- and Z-isomers of the Ostrinia pheromone. The simulation studies showed that while protein flexibility decreases upon binding to E-pheromone, it increases when bound to Z-pheromone. This finding suggests that the OnubPBP3 complex with E-pheromone is more stable than with Z-pheromone.
Subject(s)
Carrier Proteins/chemistry , Insect Proteins/chemistry , Intercellular Signaling Peptides and Proteins/chemistry , Moths , Sex Attractants , Animals , Pheromones , Zea maysABSTRACT
Asparagine-linked glycosylation, also known as N-linked glycosylation, is an essential and highly conserved co- and post-translational protein modification in eukaryotes and some prokaryotes. In the central step of this reaction, a carbohydrate moiety is transferred from a lipid-linked donor to the side-chain of a consensus asparagine in a nascent protein as it is synthesized at the ribosome. Complete loss of oligosaccharyltransferase (OST) function is lethal in eukaryotes. This reaction is carried out by a membrane-associated multisubunit enzyme, OST, localized in the endoplasmic reticulum. The smallest subunit, Ost4, contains a single membrane-spanning helix that is critical for maintaining the stability and activity of OST. Mutation of any residue from Met18 to Ile24 of Ost4 destabilizes the enzyme complex, affecting its activity. Here, we report solution nuclear magnetic resonance structures and molecular dynamics (MD) simulations of Ost4 and Ost4V23D in micelles. Our studies revealed that while the point mutation did not impact the structure of the protein, it affected its position and solvent exposure in the membrane mimetic environment. Furthermore, our MD simulations of the membrane-bound OST complex containing either WT or V23D mutant demonstrated disruption of most hydrophobic helix-helix interactions between Ost4V23D and transmembrane TM12 and TM13 of Stt3. This disengagement of Ost4V23D from the OST complex led to solvent exposure of the D23 residue in the hydrophobic pocket created by these interactions. Our study not only solves the structures of yeast Ost4 subunit and its mutant but also provides a basis for the destabilization of the OST complex and reduced OST activity.
Subject(s)
Hexosyltransferases , Saccharomyces cerevisiae Proteins , Hexosyltransferases/metabolism , Magnetic Resonance Spectroscopy , Membrane Proteins , Molecular Dynamics Simulation , Mutation , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
Herein, we report a novel, low-temperature solvothermal method to grow 3D-Bi2O3 flower-like microspheres on Ti substrates as a binder-free negative electrode for supercapacitor applications. The Bi2O3/Ti electrode showed an areal capacitance of 1.65 F cm-2 at 4 mA cm-2. Moreover, the 3D-NiCo2O4||3D-Bi2O3 hybrid device delivered high energy and power densities of 31.17 µW h cm-2 and 7500 µW cm-2, respectively. The more optimal energy storage performance based on the strong adhesion of the current collector and self-assembled three-dimensional nanostructures permits efficient electron and ion transportation.
ABSTRACT
Asparagine-linked glycosylation, also known as N-linked glycosylation is an essential and highly conserved post-translational protein modification that occurs in all three domains of life. This modification is essential for specific molecular recognition, protein folding, sorting in the endoplasmic reticulum, cell-cell communication, and stability. Defects in N-linked glycosylation results in a class of inherited diseases known as congenital disorders of glycosylation (CDG). N-linked glycosylation occurs in the endoplasmic reticulum (ER) lumen by a membrane associated enzyme complex called the oligosaccharyltransferase (OST). In the central step of this reaction, an oligosaccharide group is transferred from a lipid-linked dolichol pyrophosphate donor to the acceptor substrate, the side chain of a specific asparagine residue of a newly synthesized protein. The prokaryotic OST enzyme consists of a single polypeptide chain, also known as single subunit OST or ssOST. In contrast, the eukaryotic OST is a complex of multiple non-identical subunits. In this review, we will discuss the biochemical and structural characterization of the prokaryotic, yeast, and mammalian OST enzymes. This review explains the most recent high-resolution structures of OST determined thus far and the mechanistic implication of N-linked glycosylation throughout all domains of life. It has been shown that the ssOST enzyme, AglB protein of the archaeon Archaeoglobus fulgidus, and the PglB protein of the bacterium Campylobactor lari are structurally and functionally similar to the catalytic Stt3 subunit of the eukaryotic OST enzyme complex. Yeast OST enzyme complex contains a single Stt3 subunit, whereas the human OST complex is formed with either STT3A or STT3B, two paralogues of Stt3. Both human OST complexes, OST-A (with STT3A) and OST-B (containing STT3B), are involved in the N-linked glycosylation of proteins in the ER. The cryo-EM structures of both human OST-A and OST-B complexes were reported recently. An acceptor peptide and a donor substrate (dolichylphosphate) were observed to be bound to the OST-B complex whereas only dolichylphosphate was bound to the OST-A complex suggesting disparate affinities of two OST complexes for the acceptor substrates. However, we still lack an understanding of the independent role of each eukaryotic OST subunit in N-linked glycosylation or in the stabilization of the enzyme complex. Discerning the role of each subunit through structure and function studies will potentially reveal the mechanistic details of N-linked glycosylation in higher organisms. Thus, getting an insight into the requirement of multiple non-identical subunits in the N-linked glycosylation process in eukaryotes poses an important future goal.
Subject(s)
Hexosyltransferases/chemistry , Hexosyltransferases/metabolism , Membrane Proteins/chemistry , Membrane Proteins/metabolism , Amino Acid Sequence , Animals , Catalytic Domain , Glycosylation , Humans , Isoenzymes/chemistry , Isoenzymes/metabolism , Models, Molecular , Protein Subunits/chemistry , Protein Subunits/metabolismABSTRACT
Asparagine-linked glycosylation is an essential and highly conserved protein modification reaction that occurs in the endoplasmic reticulum of cells during protein synthesis at the ribosome. In the central reaction, a pre-assembled high-mannose sugar is transferred from a lipid-linked donor substrate to the side-chain of an asparagine residue in an -N-X-T/S- sequence (where X is any residue except proline). This reaction is carried by a membrane-bound multi-subunit enzyme complex, oligosaccharyltransferase (OST). In humans, genetic defects in OST lead to a group of rare metabolic diseases collectively known as Congenital Disorders of Glycosylation. Certain mutations are lethal for all organisms. In yeast, the OST is composed of nine non-identical protein subunits. The functional enzyme complex contains eight subunits with either Ost3 or Ost6 at any given time. Ost4, an unusually small protein, plays a very important role in the stabilization of the OST complex. It bridges the catalytic subunit Stt3 with Ost3 (or Ost6) in the Stt3-Ost4-Ost3 (or Ost6) sub-complex. Mutation of any residue from M18-I24 in the trans-membrane helix of yeast Ost4 negatively impacts N-linked glycosylation and the growth of yeast. Indeed, mutation of valine23 to an aspartate impairs OST function in vivo resulting in a lethal phenotype in yeast. To understand the structural mechanism of Ost4 in the stabilization of the enzyme complex, we have initiated a detailed investigation of Ost4 and its functionally important mutant, Ost4V23D. Here, we report the backbone 1H, 13C, and 15N resonance assignments for Ost4 and Ost4V23D in dodecylphosphocholine micelles.
Subject(s)
Carbon-13 Magnetic Resonance Spectroscopy , Hexosyltransferases/chemistry , Membrane Proteins/chemistry , Mutant Proteins/chemistry , Protein Subunits/chemistry , Proton Magnetic Resonance Spectroscopy , Saccharomyces cerevisiae/enzymology , Amino Acid Sequence , Nitrogen Isotopes , Protein Structure, SecondarySubject(s)
Lymphocytes , Neutrophils , Cerebral Hemorrhage , Humans , Prognosis , Retrospective StudiesABSTRACT
The recycling of polyvinyl chloride (PVC) recovered from the plastic insulations in wires and cables is a rising concern in the current situation due to its hazardous behaviour during recycling. Similarly, high-impact polystyrene (HIPS) and acrylonitrile butadiene styrene (ABS) used in the structural components of electrical and electronic equipment are also generated in large quantities. In the current work, three agendas were fixed: (a) to determine the effect of recycled polymeric material (HIPS and ABS) recovered from different sources on the mechanical property of the polymeric blends; (b) to formulate a high-impact strength blend; and (c) to deduce a mechanism for improved impact strength. The mechanical characterizations were conducted on the entire blends formulated. Among them, the recycled blend composed of recycled PVC (r-PVC) and recycled ABS (r-ABS) (segregated from uninterrupted power supply housing) and recycled HIPS (r-HIPS; collected from television housing) was confined for further physio-mechanical and thermal analysis. Besides, the r-PVC/r-ABS systems had shown better mechanical properties than r-PVC/r-HIPS systems in similar composition. The impact strength of blend r-PVC/r-ABS (70:30) was found to be 250 J/m, which was 200% more than the blend r-PVC/r-ABS (0:100). The compatibility and non-compatibility in PVC/ABS and PVC/HIPS blends respectively were explained with thermal, mechanical and morphological characterizations. Furthermore, a plausible cross-linking mechanism is developed between ABS and PVC, which controls the release of chlorine atoms into the environment.
Subject(s)
Acrylonitrile , Electronic Waste , Hazardous Waste , Plastics , RecyclingABSTRACT
Epoxy-polydimethylsiloxane-graphene oxide (EPG) nanocomposite coatings were successfully developed by loading different wt% of graphene oxide nanosheets (GNs) into an epoxy-hydroxy-terminated-polydimethylsiloxane (EP-hPD) matrix via a facile in situ preparation technique. The inclusion of GNs into EPN led to an increase in modulus of elasticity and tensile strength up to 1570.46 MPa and 31.54 MPa, respectively, in the case of 1 wt% loading of GNs in the EP-hPD matrix. Also, an increase in the water contact angle from 90.1° to 115.2°, 104.5° and 101.7° was discerned at 1, 3 and 5 wt% loadings of GNs respectively. Taber abrasion results demonstrated a decrease in abrasion loss by 33.3% at 1 wt% loading of GNs in comparison to the unreinforced coating. An improvement in the glass transition temperature (Tg) was observed from 63.5 °C for the neat sample to 77.6 °C, 76.3 °C and 71.6 °C for the 1, 3 and 5 wt% EPG nanocomposites, respectively. An inevitable enhancement in the properties of the nanocomposites was affirmed due to the synergistic effect of GNs dispersed within the EP-hPD blend matrix. The prominent findings of this work include a minimum corrosion rate of 0.73 × 10-2 mm per year and upgradation in the antifouling performance of the nanocomposite coatings in comparison to the neat coating.
ABSTRACT
Ostrinia furnacalis, a lepidopteran moth, is an invasive pest found in Asia, Australia, Africa, and parts of the United States. The O. furnacalis pheromone-binding protein2 (OfurPBP2), present in the male moth antenna, plays a role in the detection of female-secreted pheromone in a process that leads to mating. To understand the structural mechanism of pheromone binding and release in this pest, we have initiated characterization of OfurPBP2 by solution NMR. Here, we report the backbone resonance assignments and the secondary structural elements of OfurPBP2 at pH 6.5 using uniformly 13C, 15N-labeled protein with various triple resonance NMR experiments. The assignments are 97% completed for backbone and 88% completed for side-chain resonances. The secondary structure of OfurPBP2, based on backbone chemical shifts, consists of eight α-helices, including a well-structured C-terminal helix.
Subject(s)
Agriculture , Carbon-13 Magnetic Resonance Spectroscopy , Carrier Proteins/chemistry , Insect Proteins/chemistry , Moths/metabolism , Nuclear Magnetic Resonance, Biomolecular , Proton Magnetic Resonance Spectroscopy , Animals , Nitrogen Isotopes , Protein Structure, SecondaryABSTRACT
Pheromone-binding protein (PBP) in male moth antennae transports pheromone to the olfactory receptor neuron by undergoing a pH-dependent conformational switch, from PBPB at higher pH to PBPA at lower pH, associated with ligand binding and release, respectively. The characteristic feature of the dramatic protein switch is the pH-dependent reversible coil-helix transition of the C-terminus. In the PBPB conformation at pH >6.0, the C-terminus is exposed to the solvent as a coil while the ligand occupies the hydrophobic pocket. However, in the PBPA conformation at acidic pH, the C-terminus switches to a helix and releases the ligand by outcompeting it for the hydrophobic pocket. In Antheraea polyphemus PBP1 (ApolPBP1), the C-terminus (P129-V142) is composed predominantly of hydrophobic residues except for three strategically located acidic residues: Asp132, Glu137, and Glu141. Here, we report for the first time on the consequences of the mutation of one or more acidic residues in the pH-driven reversible coil-helix transition of the ApolPBP1 C-terminus through biophysical characterization. Mutation of any single acidic residue in the C-terminus to its neutral counterpart destabilizes the helix formation at lower pH; these mutants exist as a mixture of both conformations. However, mutation of the two terminal acidic residues together knocks out the protein switch and adversely affects both ligand binding and release functions. Thus, these mutant proteins remain in the open (PBPB) conformation at all pH levels.