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Fibrillar aggregates of the protein α-synuclein (αS) are one of the hallmarks of Parkinson’s disease. Here, we show that measuring the fluorescence polarization (FP) of labels at several sites on αS allows one to monitor changes in the local dynamics of the protein after binding to micelles or vesicles, and during fibril formation. Most significantly, these site-specific FP measurements provide insight into structural remodeling of αS fibrils by small molecules and have the potential for use in moderate-throughput screens to identify small molecules that could be used to treat Parkinson’s disease. © 2016 American Chemical Society.

Ammonia in the tetrazole Ugi variation together with α-amino acid methyl ester-derived isocyanides provides tetrazolopiperidinones in good to high yields in one pot. The scope and limitations of this reaction were investigated by performing >70 reactions. The scaffold is useful to fill high-throughput screening decks and in structure-based drug design.

Despite the high prevalence of neuropsychiatric disorders, their aetiology and molecular mechanisms remain poorly understood. The zebrafish (Danio rerio) is increasingly utilized as a powerful animal model in neuropharmacology research and in vivo drug screening. Collectively, this makes zebrafish a useful tool for drug discovery and the identification of disordered molecular pathways. Here, we discuss zebrafish models of selected human neuropsychiatric disorders and drug-induced phenotypes. As well as covering a broad range of brain disorders (from anxiety and psychoses to neurodegeneration), we also summarize recent developments in zebrafish genetics and small molecule screening, which markedly enhance the disease modelling and the discovery of novel drug targets. © 2017 The British Pharmacological Society The study was coordinated through the International Zebrafish Neuroscience Research Consortium (ZNRC), and this collaboration was funded by St. Petersburg State University, Ural Federal University and Guangdong Ocean University. A.V.K. is the Chair of ZNRC, and his research is supported by the Russian Foundation for Basic Research (RFBR) grant 16-04-00851.

LC/MS methodology for the analysis of domoic acidand lipophilic toxins in shellfish was developed using a hybrid triple quadrupole linear ion trap mass spectrometer. For routine quantitation a scheduled selected reaction monitoring (SRM) method for the analysis of domoic acid, okadaic acid, dinophysistoxins,azaspiracids, pectenotoxins, yessotoxins, gymnodimines, spirolides, and pinnatoxins was developed and validated. The method performed well in terms of LOD, linearity, precision, and trueness. Taking advantageof the high instrument sensitivity, matrix effects were mitigated by reducing the amount of sample introduced to the mass spectrometer. Optionally, samples can be analyzed using information dependent acquisition (IDA) methods, either in positive or negative mode, which can provide an extra level of confirmationby matching the full product ion spectra acquired for a sample with those from a specially constructedspectral library. Methods were applied to the analysisof a new certified reference material and Canadian mussels (Mytilus edulis) implicated in a 2011 diarrhetic shellfish poisoning (DSP) incident. The scheduled SRM method enabled the screening and quantitation of multiple phycotoxins. As DSPhad not previously been observed in this area of Canada,positive identification of putative toxins was accomplished using the IDA and spectral search method. Analysis of the 2011 toxic mussel samples revealed thepresence of high levels of dinophysistoxin-1, which explained the DSP symptoms, as well as pectenotoxins, yessotoxins, and variety of cyclic imines.

The genetic missense A30P mutation of the wild-type α-synuclein protein results in the replacement of the 30th amino acid residue from alanine (Ala) to proline (Pro) and was initially found in the members of a German family who developed Parkinson's disease. Even though the structures of these proteins have been measured before, detailed understanding about the structures and their relationships with free energy landscapes is lacking, which is of interest to provide insights into the pathogenic mechanism of Parkinson's disease. We report the secondary and tertiary structures and conformational free energy landscapes of the wild-type and A30P mutant-type α-synuclein proteins in an aqueous solution environment via extensive parallel tempering molecular dynamics simulations along with thermodynamic calculations. In addition, we present the residual secondary structure component transition stabilities at the atomic level with dynamics in terms of free energy change calculations using a new strategy that we reported most recently. Our studies yield new interesting results; for instance, we find that the A30P mutation has local as well as long-range effects on the structural properties of the wild-type α-synuclein protein. The helical content at Ala18-Gly31 is less prominent in comparison to the wild-type α-synuclein protein. The β-sheet structure abundance decreases in the N-terminal region upon A30P mutation of the wild-type α-synuclein, whereas the NAC and C-terminal regions possess larger tendencies for β-sheet structure formation. Long-range intramolecular protein interactions are less abundant upon A30P mutation, especially between the NAC and C-terminal regions, which is linked to the less compact and less stable structures of the A30P mutant-type rather than the wild-type α-synuclein protein. Results including the usage of our new strategy for secondary structure transition stabilities show that the A30P mutant-type α-synuclein tendency toward aggregation is higher than the wild-type α-synuclein but we also find that the C-terminal and NAC regions of the A30P mutant-type α-synuclein are reactive toward fibrillzation and aggregation based on atomic level studies with dynamics in an aqueous solution environment. Therefore, we propose that small molecules or drugs blocking the specific residues, which we report herein, located in the NAC- and C-terminal regions of the A30P mutant-type α-synuclein protein might help to reduce the toxicity of the A30P mutant-type α-synuclein protein. © 2013 American Chemical Society.

A complete library of new alkoxides containing group 4 metals and the imino phenol ligand were synthesized in high yields and purity. These compounds showed catalytic activity towards the polymerization of l-lactide (l-LA), rac-lactide (rac-LA), ?-caprolactone (CL), ?-valerolactone (VL), rac-?-butyrolactone (rac-BL), ethylene and propylene. The zirconium catalysts were found to yield better polymerization results in comparison to the titanium and hafnium analogues. The number average molecular weight (M n) values of the polymers are very high for LA and CL with controlled molecular weight distributions (MWDs). Analyses of MALDI-TOF and 1H NMR spectra of low Mn oligomers reveal that the ligand initiates the ring-opening polymerization (ROP). For ethylene and propylene polymerization, we have achieved moderate to good activity using MAO as a co-catalyst. � 2013 The Royal Society of Chemistry and the Centre National de la Recherche Scientifique.

Single-domain antibodies (sdAbs) derived from human V H are considered to be less soluble and prone to aggregate which makes it difficult to determine the crystal structures. In this study, we isolated and characterized two anti-human epidermal growth factor receptor-2 (HER2) sdAbs, Gr3 and Gr6, from a synthetic human V H phage display library. Size exclusion chromatography and surface plasmon resonance analyses demonstrated that Gr3 is a monomer, but that Gr6 is a strict dimer. To understand this different molecular behavior, we solved the crystal structure of Gr6 to 1.6 Å resolution. The crystal structure revealed that the homodimer assembly of Gr6 closely mimics the V H-V L heterodimer of immunoglobulin variable domains and the dimerization interface is dominated by hydrophobic interactions. © 2012 Baral et al.

The ability of a peptide to target a specific protein in vitro has a potential for recognizing the cell membrane structure, protein-protein and receptor-ligand interaction. On the basis of molecular interactions, cell specific peptides were selected via phage library approach and their functions on the cells were determined by XTT based viability assay. We aimed to find phage displayed peptides from 12-mer peptide library that interact with K562-dox cell membrane in association with cellular functions and to study the effects of peptides selected from artificial peptide library on K562-dox cell viability. Peptides recognizing K562-dox cells were identified according to peptide sequences and amino acid properties. We selected 29 different phages from biopannings with K562-dox cells. Three clones were identified (KPB7,KPB10, KPP8) and their negative effects on cell viability were determined by XTT assay. According to our cell viability assay results, selected phages were effected negatively to the viability of the K562-dox cells. Depending on the present results, peptides were obtained that could be potential candidates for molecular recognition and cell targeting approaches. © 2010 Academic Journals Inc.