Interaction of Synthetic Human SLURP-1 with the Nicotinic Acetylcholine Receptors

• DOI: 10.1038/s41598-017-16809-0
• Publication Date: 2017-11-24T10:19:24Z
• Authors: Thomas Durek; Irina V. Shelukhina; Han-Shen Tae; Panumart Thongyoo; Ekaterina N. Spirova; Denis S. Kudryavtsev; Igor E. Kasheverov; Grazyna Faure; Pierre-Jean Corri...; David J. Craik; David J. Adams; Victor I. Tsetlin
• Keywords: System; Models, Molecular; 0301 basic medicine; Spectrometry, Mass, Electrospray Ionization; Magnetic Resonance Spectroscopy; 572; Protein Conformation; Native Chemical Ligation; [SDV]Life Sciences [q-bio]; Neurotoxins; Modulator; Expression; Receptors, Nicotinic; Ligands; Article; Structure-Activity Relationship; 03 medical and health sciences; Allosteric Regulation; Medicine and Health Sciences; [CHIM]Chemical Sciences; Antigens, Ly; Humans; Amino Acid Sequence; Chromatography, High Pressure Liquid; Neurons; Lynx1; Muscles; Phenylurea Compounds; Proteins; Isoxazoles; Binding; Urokinase-Type Plasminogen Activator; 1000 General; Peptide; Protein Multimerization; Protein Binding

Human SLURP-1 is a secreted protein of the Ly6/uPAR/three-finger neurotoxin family that co-localizes with nicotinic acetylcholine receptors (nAChRs) and modulates their functions. Conflicting biological activities at various nAChR subtypes have been based on heterologously produced containing N- and/or C-terminal extensions. Here, we report chemical synthesis 81 amino acid residue human protein, characterization its 3D structure by NMR, activity subtypes. Radioligand assays indicated synthetic did not compete [125I]-α-bungarotoxin (α-Bgt) binding to neuronal α7 Torpedo californica muscle-type nAChRs, nor mollusk proteins (AChBP). Inhibition α7-mediated currents only occurred in presence allosteric modulator PNU120596. In contrast, observed robust mediated inhibition α3β4, α4β4, α3β2 as well rat α9α10 nAChRs. nAChRs was accentuated higher ACh concentrations, indicating an mechanism. Our results are discussed context recent studies indicate N-terminal extensions may affect selectivity targets. this respect, appears be better probe for structure-function studies.