Antagonistic Mechanism of α-Conotoxin BuIA toward the Human α3ß2 Nicotinic Acetylcholine Receptor.

Nicotinic acetylcholine receptors (nAChRs) are pentameric ligand-gated ion channels that are abundantly expressed in the central and peripheral nervous systems, playing an important role in mediating neurotransmitter release and inter-synaptic signaling. Dysfunctional nAChRs are associated with neurological disorders, and studying the structure and function of nAChRs is essential for development of drugs or strategies for treatment of related diseases. α-Conotoxins are selective antagonists of the nAChR and are an important class of drug leads. So far, the antagonistic mechanism of α-conotoxins toward the nAChRs is still unclear. In this study, we built an α3ß2 nAChR homology model and investigated its conformational transition mechanism upon binding with a highly potent inhibitor, α-conotoxin BuIA, through µs molecular dynamic simulations and site-directed mutagenesis studies. The results suggested that the α3ß2 nAChR underwent global conformational transitions and was stabilized into a closed state with three hydrophobic gates present in the transmembrane domain by BuIA. Finally, the probable antagonistic mechanism of BuIA was proposed. Overall, the closed-state model of the α3ß2 nAChR bound with BuIA is not only essential for understanding the antagonistic mechanism of α-conotoxins but also particularly valuable for development of therapeutic inhibitors in future.

Chitin nanofibers as versatile bio-templates of zeolitic imidazolate frameworks for N-doped hierarchically porous carbon electrodes for supercapacitor.

Biobased N-doped hierarchically porous carbon (N-HPC) electrodes were successfully prepared by utilizing marine crustacean derivatives and chitin nanofibers (ChNF), as versatile bio-templates of zeolitic imidazolate frameworks (ZIF-8) to form ChNF@ZIF-8 nanocomposites, followed by a subsequent carbonization process. The ZIF-8 nanoparticles were in situ synthesized on ChNF surfaces to avoid fragmentation for fabricating hierarchically porous carbon structure (N-HPC), which is efficiently doped with rich nitrogen content that originates in ChNF and ZIF-8. The results show that N-HPC electrodes demonstrate improved electrochemical performance and the constructed symmetric supercapacitor assembled with N-HPC exhibits enhanced capacitive performance of specific capacity (128.5 F·g-1 at 0.2 A·g-1) and excellent electrochemical stability even after 5000 cycles. This facile and effective preparation method of N-HPC electrodes derived from marine crustacean nanomaterials will have great potential in the construction of next-generation electrochemical energy-storage devices with excellent capacitance performance.

Exploring the Antihyperglycemic Chemical Composition and Mechanisms of Tea Using Molecular Docking.

Tea, a widely consumed beverage, has long been utilized for promoting human health with a close correlation to hyperglycemia. The Tea Metabolome Database (TMDB), the most complete and comprehensive curated collection of tea compounds data containing 1271 identified small molecule compounds from the tea plant (Camellia sinensis), was established previously by our research team. More recently, our studies have found that various tea types possess an antihyperglycemic effect in mice. However, the bioactive ingredients from tea have potential antihyperglycemic activity and their underlying molecular mechanisms remain unclear. In this study, we used a molecular docking approach to investigate the potential interactions between a selected 747 constituents contained in tea and 11 key protein targets of clinical antihyperglycemic drugs. According to our results, the main antihyperglycemic targets of tea composition were consistent with those of the drug rosiglitazone. The screening results showed that GCG, ECG3'Me, TMDB-01443, and CG had great target binding capacity. The results indicated that these chemicals of tea might affect hyperglycemia by acting on protein targets of rosiglitazone.

Cellulose, hemicellulose, lignin, and their derivatives as multi-components of bio-based feedstocks for 3D printing.

Three-dimensional (3D) printing, known as revolutionary and disruptive innovation in manufacturing technology, supports great opportunities to rapidly construct a wide range of tailored object geometries. Cellulose, hemicellulose, and lignin as the three most common natural polymers and main components of plant resources, possess great economical potential for bio-based products due to their attractive advantages. The integration of 3D printing technology involved with cellulose, hemicellulose and lignin as the major bio-based feedstock for high-performance 3D printed products has received great concern in the R&D areas. In this review, the aim is to shed light on a cutting-edge review on the most recent progress based on cellulose, hemicellulose and lignin, as well as their derivatives as multi-components of bio-feedstock for 3D printing, in which the applications, roles and functions of the plant-derived biomass for 3D printing are also highlighted. The challenges and perspectives for future work are provided, to underscore critical issues and opportunities.

Rational Design of α-Conotoxin RegIIA Analogues Selectively Inhibiting the Human α3ß2 Nicotinic Acetylcholine Receptor through Computational Scanning.

Engineering the selectivity of α-conotoxins for nicotinic acetylcholine receptors (nAChRs) presents considerable complexity and challenges, as it involves the optimization of their binding affinities to multiple highly conserved nAChR subtypes. Here, we applied a computational-based scanning approach for the rational design of α-conotoxin RegIIA analogues selectively targeting the human (h) α3ß2 versus hα3ß4 nAChRs. Binding mode analyses suggested that several residues in loop II of RegIIA (position 9, 10, and 11) formed nonconserved interactions with residues of the ß2 and ß4 subunits. The molecular mechanics generalized Born surface area method was applied for in silico sequence scanning of RegIIA position 9, 10, and 11 on frames extracted from single molecular dynamics simulation trajectory. RegIIA analogues with favorable predicted binding affinities solely to the hα3ß2 nAChR were synthesized and tested electrophysiologically. We report three RegIIA analogues, with position 9 aromatic residue substitutions, exhibiting a 10- to 37-fold subtype selectivity improvement for hα3ß2 compared to hα3ß4 nAChR. The in silico scanning method proposed from this study has considerable potential for the efficient design of nAChR subtype selective antagonists in the future.

Ball milling pretreatment facilitating α-amylase hydrolysis for production of starch-based bio-latex with high performance.

Starch based bio-latex has been widely researched in the coating paper area for the purpose of partial replacement of petroleum-based binders. In this paper, a green and facile ball milling pretreatment was proposed to modify the starch granules before α-amylase hydrolysis by breaking up their crystalline structure, thus improving the accessibility and susceptibility of amylase into starch structure. It was found that the improved hydrolysis process after 8 h ball milling can generate suitable degree of polymerization of polysaccharides or oligosaccharides, which further facilitated the following H2O2 oxidation and SHMP crosslinking processes. In addition, a mechanism was also demonstrated to illustrate the improvement induced by ball milling pretreatment. The prepared bio-latex with crosslinking-structure performed excellent adhesive properties when substituted 25 % of petroleum-based latex during paper coating application, which showed great potential in improving the economic, cost, and environment benefits of traditional production of coated paper.

Preparation and characterization of high yield cellulose nanocrystals (CNC) derived from ball mill pretreatment and maleic acid hydrolysis.

The target of the study is to improve the yield and the colloidal stability of cellulose nano-crystals (CNC) that is obtained through maleic acid hydrolysis. Herein, a facile/ green approach to prepare CNC with high yield and colloidal stability from bamboo fibers is presented. Ball mill pretreatment can break down and open up the structure of bamboo fibers, thus exposing more hydroxyl groups on the surface of pulp fibers and increasing the access of acid molecules into pulp fibers. The maleic acid molecules can easily hydrolyze cellulose, thus releasing more crystalline parts; maleic acid anhydride can react with hydroxyl groups to generate more -COOH groups on CNC. The yield of resultant CNC was 10.55-24.50 %, which was much higher than 2.80 % of the control. The study put forward a facile approach to prepare CNC with high yield and colloidal stability, and paves a possible way for industrialization of CNC production.

Enhanced Activity against Multidrug-Resistant Bacteria through Coapplication of an Analogue of Tachyplesin I and an Inhibitor of the QseC/B Signaling Pathway.

Tachyplesin I (TPI) is a cationic ß-hairpin antimicrobial peptide with broad-spectrum, potent antimicrobial activity. In this study, the all d-amino acid analogue of TPI (TPAD) was synthesized, and its structure and activity were determined. TPAD has comparable antibacterial activity to TPI on 14 bacterial strains, including four drug-resistant bacteria. Importantly, TPAD has significantly improved stability against enzymatic degradation and decreased hemolytic activity compared to TPI, indicating that it has better therapeutic potential. The induction of bacterial resistance using low concentrations of TPAD resulted in the activation of the QseC/B two-component system. Deletion of this system resulted in at least five-fold improvement of TPAD activity, and the combined use of TPAD with LED209, a QseC/B inhibitor, significantly enhanced the bactericidal effect against three classes of multidrug-resistant bacteria.

In Silico Design of MDM2-Targeting Peptides from a Naturally Occurring Constrained Peptide.

Naturally occurring constrained peptides are frequently used as scaffolds for bioactive peptide grating due to their high stability. Here, we used in silico methods to design several constrained peptides comprising a scorpion toxin scaffold, a MDM2 binding epitope, and a cluster of positively charged residues. The designed peptides displayed varied binding affinity to MDM2 despite differing by only one or two residues. One of the peptides, SC426, had nanomolar binding affinity (KD =6.6±2.6 nm) to MDM2, and exhibited stronger inhibitory activity on the proliferation of HCT116 cells (p53-wild type) and SW480 cells (p53-mutant) than that of nutlin-3a. Binding mode analysis of the designed peptide at MDM2 suggests that the conserved "FWL" epitope was buried in the hydrophobic binding pocket, and the residues located at the periphery of the binding site contributed to the high binding affinity of SC426. Overall, in silico design of miniproteins with therapeutic potential through epitope grafting to the naturally occurring constrained peptide is an effective strategy.

α-Conotoxin Vc1.1 Structure-Activity Relationship at the Human α9α10 Nicotinic Acetylcholine Receptor Investigated by Minimal Side Chain Replacement.

α-Conotoxin Vc1.1 inhibits the nicotinic acetylcholine receptor (nAChR) α9α10 subtype and has the potential to treat neuropathic chronic pain. To date, the crystal structure of Vc1.1-bound α9α10 nAChR remains unavailable; thus, understanding the structure-activity relationship of Vc1.1 with the α9α10 nAChR remains challenging. In this study, the Vc1.1 side chains were minimally modified to avoid introducing large local conformation perturbation to the interactions between Vc1.1 and α9α10 nAChR. The results suggest that the hydroxyl group of Vc1.1, Y10, forms a hydrogen bond with the carbonyl group of α9 N107 and a hydrogen bond donor is required. However, Vc1.1 S4 is adjacent to the α9 D166 and D169, and a positive charge residue at this position increases the binding affinity of Vc1.1. Furthermore, the carboxyl group of Vc1.1, D11, forms two hydrogen bonds with α9 N154 and R81, respectively, whereas introducing an extra carboxyl group at this position significantly decreases the potency of Vc1.1. Second-generation mutants of Vc1.1 [S4 Dab, N9A] and [S4 Dab, N9W] increased potency at the α9α10 nAChR by 20-fold compared with that of Vc1.1. The [S4 Dab, N9W] mutational effects at positions 4 and 9 of Vc1.1 are not cumulative but are coupled with each other. Overall, our findings provide valuable insights into the structure-activity relationship of Vc1.1 with the α9α10 nAChR and will contribute to further development of more potent and specific Vc1.1 analogues.

Molecular dynamics simulations of dihydro-ß-erythroidine bound to the human α4ß2 nicotinic acetylcholine receptor.

BACKGROUND AND PURPOSE: The heteromeric α4ß2 nicotinic acetylcholine receptor (nAChR) is abundant in the human brain and is associated with a range of CNS disorders. This nAChR subtype has been recently crystallised in a conformation that was proposed to represent a desensitised state. Here, we investigated the conformational transition mechanism of this nAChR from a desensitised to a closed/resting state. EXPERIMENTAL APPROACH: The competitive antagonist dihydro-ß-erythroidine (DHßE) was modelled by replacement of the agonist nicotine in the α4ß2 nAChR experimental structure. DHßE is used both in vitro and in vivo for its ability to block α4ß2 nAChRs. This system was studied by three molecular dynamics simulations with a combined simulation time of 2.6 µs. Electrophysiological studies of mutated receptors were performed to validate the simulation results. KEY RESULTS: The relative positions of the extracellular and transmembrane domains in the models are distinct from those of the desensitised state structure and are compatible with experimental structures of Cys-loop receptors captured in a closed/resting state. CONCLUSIONS AND IMPLICATIONS: Our model suggests that the side chains of α4 L257 (9') and α4 L264 (16') are the main constrictions in the transmembrane pore. The involvement of position 9' in channel gating is well established, but position 16' was only previously identified as a gate for the bacterial channels, ELIC and GLIC. L257 but not L264 was found to influence the slow component of desensitisation. The structure of the antagonist-bound state proposed here should be valuable for the development of therapeutic or insecticide compounds.

Role of the disulfide bond on the structure and activity of µ-conotoxin PIIIA in the inhibition of NaV1.4.

µ-Conotoxin PIIIA, a peptide toxin isolated from Conus purpurascens, blocks the skeletal muscle voltage-gated sodium channel NaV1.4 with significant potency. PIIIA has three disulfide bonds, which contribute largely to its highly constrained and stable structure. In this study, a combination of experimental studies and computational modeling were performed to assess the effects of deletion of the disulfide bonds on the structure and activity of PIIIA. The final results indicate that the three disulfide bonds of PIIIA are required to produce the effective inhibition of NaV1.4, and the removal of any one of the disulfide bonds significantly reduces its binding affinity owing to secondary structure variation, among which the Cys11-Cys22 is the most important for sustaining the structure and activity of PIIIA.

Synthesis of novel 10,11-methylenedioxy-camptothecin glycoside derivatives and investigation of their anti-tumor effects in vivo.

10,11-Methylenedioxy-camptothecin (FL118) is a novel camptothecin analogue that possesses exceptional antitumor efficacy in human tumor xenograft models. The aim of the current study was to develop novel 20-substituted FL118 derivatives coupled with glycosyl-succinic acid esters with improved antitumor efficacy. These FL118 glycoside derivatives were designed, synthesized and their cytotoxicity evaluated in three tumor cell lines (A-549, MDA-MB-231 and RM-1). All of the derivatives showed superior in vitro cytotoxic activity and were more potent than irinotecan in A549 and MDA-MB-231 cells. In mouse prostate cancer cells RM-1, 10,11-methylenedioxy-camptothecin rhamnoside 11b displayed significant activities with IC50 of 48.27 nM. Western blot analysis demonstrated that 11b inhibited survivin expression and induced cancer cells apoptosis. Further cell cycle analyses clearly showed 11b induced G2/M phase cell cycle arrest. Molecule docking studies suggested that the binding mode of 11b was different from that of the crystal complex of ligand topotecan in Top1/DNA. Importantly, 11b showed high in vivo antitumor efficacy in the RM-1 mouse model with transplantation of prostate cancer (TGI = 44.9%) at dose of 9 mg kg-1 without apparent toxicity.

Application of reverse docking for target prediction of marine compounds with anti-tumor activity.

A large number of structures of anti-cancer drug targets have been solved and deposited to the protein data bank already. Identification of the targets for marine compounds with anti-tumor activity presents a challenge for marine natural products scientists. In this study, fast and efficient computational reverse docking was applied to predict the probable targeting proteins of the marine compounds with anti-tumor activity. Crystal structures of the proteins involved in tumor genesis, growth and metastasis were collected from PDB to construct the anti-tumor protein database (APD) for reverse docking. Two non-commercial docking programs, AutoDock Vina and LeDock, were used to perform the docking. Our results suggest that reverse docking is efficient for target fishing of compounds with known anti-tumor activities. In addition, the results show that performance of reverse docking using LeDock is superior to that using AutoDock Vina. Overall, reverse docking is a fast and efficient computational method to identify the probable target of the compounds with anti-tumor activities, and it can be complementary to the biological testing methods.