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nmrlearner 08-24-2020 09:54 PM

Solution structure of insect CSP and OBPs by NMR.
 
Solution structure of insect CSP and OBPs by NMR.

Solution structure of insect CSP and OBPs by NMR.

Methods Enzymol. 2020;642:169-192

Authors: Leone S, Emendato A, Spadaccini R, Picone D

Abstract
Insect odorant binding proteins (OBPs) and chemosensory proteins (CSPs) are proteins deputed to the solubilization, transport and stabilization of lipophilic and odorant compounds. These proteins have a conserved fold, which undergoes massive structural rearrangements in order to accommodate medium to large-sized lipophilic ligands. Solution NMR spectroscopy, due to its intrinsically dynamic nature, is the perfect technique to extrapolate structural information and dynamic parameters and to elucidate the conformational changes that occur upon ligand binding. This chapter will describe in detail the experimental protocols for the production and purification of isotope-labeled recombinant CSPs and OBPs for NMR studies. Detailed procedures for spectra acquisition, processing and analysis will be presented, focusing on the protein CSP-sg4 from Schistocerca gregaria as a model. Finally, experiments aimed at providing information on protein flexibility and ligand binding modes will also be described.


PMID: 32828252 [PubMed - as supplied by publisher]



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