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(LEXSY) system is an inexpensive and effective expression approach for various research and medical purposes. The stated advantages of this system are the possibility of obtaining the soluble product in the cytoplasm, a high probability of correct protein folding with a full range of post-translational modifications (including uniform glycosylation), and the possibility of expressing multi-subunit proteins. In this paper, a LEXSY expression system has been employed for obtaining the receptor binding domain (RBD) of the spike-protein of the SARS-CoV-2 virus and the homopentameric acetylcholine-binding protein (AChBP) from . RBD is actively used to obtain antibodies against the virus and in various scientific studies on the molecular mechanisms of the interaction of the virus with host cell targets. AChBP represents an excellent structural model of the ligand-binding extracellular domain of all subtypes of nicotinic acetylcholine receptors (nAChRs). Both products were obtained in a soluble glycosylated form, and their structural and functional characteristics were compared with those previously described.
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http://dx.doi.org/10.3390/molecules29050943 | DOI Listing |
RSC Chem Biol
August 2025
Department of Chemistry - BMC, Uppsala University Sweden
Analysis of ligand-induced structural changes in proteins is challenging due to the lack of experimental methods suited for detection and characterisation of both ligand binding and induced structural changes. We have explored biosensors with different detection principles to study interactions between ligands and acetylcholine binding proteins (AChBPs), soluble homologues of Cys-loop ligand gated ion channels (LGICs) that undergo similar structural changes as LGICs upon ligand binding. X-ray crystallography was used to identify binding sites and establish if the detected conformational changes involved small changes in loop C or major structural changes in the pentamer associated with ion channel opening.
View Article and Find Full Text PDFPhys Chem Chem Phys
September 2025
CQC-IMS, Department of Chemistry, University of Coimbra, 3004-535 Coimbra, Portugal.
The design of new materials that can be effective for adsorbing pesticides constitutes an important contribution for water remediation. Amino acids are the building blocks of proteins that bind pesticides in living organisms, and thus, they are expected to contribute to improve the adsorption properties of materials for water remediation. In this work, we propose a multi-stage computational strategy, based on docking, molecular dynamics (MD) simulations and electronic-structure calculations, to unveil relevant interactions between pesticides and amino acids from typical target proteins.
View Article and Find Full Text PDFInt Rev Neurobiol
July 2025
Medical Sciences Division, Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, United Kingdom. Electronic address:
Autoimmune Myasthenia Gravis (MG) is a disease characterized by fatigable muscle weakness and autoantibodies. It can be divided by the presence of serum autoantibodies into two major categories where Immunoglobulin G (IgG) against either the acetylcholine receptor (AChR), or muscle specific kinase (MuSK) causes fatigable muscle weakness. The clinical relevance of Low-density lipoprotein-receptor related protein-4 IgG (LRP4) is debated.
View Article and Find Full Text PDFNpj Viruses
June 2025
Department of Biomedical and Pharmaceutical Sciences, Idaho State University, Pocatello, ID, 83209, USA.
Herpes virus1(HSV1) is a neurotropic virus that has been linked to Alzheimer's disease. An In-silico structural homology search using α -Bgtx, identified structural homology between HSV1 gD and the nicotinic receptor neurotoxin α-Bgtx. SPR binding studies using acetylcholine binding protein from Lymnaea stagnalis, and functional two electrode voltage clamp studies of α7 nAChRs demonstrate the ability of HSV1 to interact directly with nAChRs.
View Article and Find Full Text PDFJ Am Chem Soc
April 2025
Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Showa-ku, Nagoya 466-8555, Japan.
The M muscarinic receptor (MR) is a prototypical G protein-coupled receptor (GPCR) that serves as a model system for understanding ligand recognition and GPCR activation. Here, using vibrational spectroscopy, we identify the mechanisms governing MR activation by its native agonist, acetylcholine. Combined with mutagenesis, computational chemistry, and organic synthetic chemistry, our analyses found that the precise distance between acetylcholine and Asn404, one of the amino acids constituting the ligand-binding site, is important for MR activation and that the N404Q mutant undergoes partial active state-like conformational changes.
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