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]
[NMR paper] Production of isotope-labeled proteins in insect cells for NMR.
Production of isotope-labeled proteins in insect cells for NMR.
Related Articles Production of isotope-labeled proteins in insect cells for NMR.
J Biomol NMR. 2018 Apr 23;:
Authors: Franke B, Opitz C, Isogai S, Grahl A, Delgado L, Gossert AD, Grzesiek S
Abstract
Baculovirus-infected insect cells have become a powerful tool to express recombinant proteins for structural and functional studies by NMR spectroscopy. This article provides an introduction into the insect cell/baculovirus expression system and its use for the production...
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04-26-2018 02:15 AM
Production of isotope-labeled proteins in insect cells for NMR
Production of isotope-labeled proteins in insect cells for NMR
Abstract
Baculovirus-infected insect cells have become a powerful tool to express recombinant proteins for structural and functional studies by NMR spectroscopy. This article provides an introduction into the insect cell/baculovirus expression system and its use for the production of recombinant isotope-labeled proteins. We discuss recent advances in inexpensive isotope-labeling methods using labeled algal or yeast extracts as the amino acid source and give examples of advanced NMR...
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04-23-2018 05:00 PM
An improved protocol for amino acid type-selective isotope labeling in insect cells
An improved protocol for amino acid type-selective isotope labeling in insect cells
Abstract
An improved expression protocol is proposed for amino acid type-specific , -isotope labeling of proteins in baculovirus-infected (BV) insect cell cultures. This new protocol modifies the methods published by Gossert et al. (J Biomol NMR 51(4):449â??456, 2011) and provides efficient incorporation of isotopically labeled amino acids, with similar yields per L versus unlabeled expression in rich media. Gossert et al. identified the presence of unlabeled amino...
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07-15-2017 05:05 PM
Insect CellGlycosylation and Its Impact on the Functionalityof a Recombinant Intracrystalline Nacre Protein, AP24
Insect CellGlycosylation and Its Impact on the Functionalityof a Recombinant Intracrystalline Nacre Protein, AP24
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.5b01186/20160208/images/medium/bi-2015-01186y_0006.gif
Biochemistry
DOI: 10.1021/acs.biochem.5b01186
http://feeds.feedburner.com/~ff/acs/bichaw?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/bichaw/~4/wwDLuPYmmFg
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02-09-2016 04:21 AM
Affordable uniform isotope labeling with 2 H, 13 C and 15 N in insect cells
Affordable uniform isotope labeling with 2 H, 13 C and 15 N in insect cells
Abstract
For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80Â*% can be achieved for 15N and 13C with yields comparable to expression in...
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04-30-2015 09:13 PM
The insect defensin lucifensin from Lucilia sericata
The insect defensin lucifensin from Lucilia sericata
The insect defensin lucifensin from Lucilia sericata
Content Type Journal Article
Category NMR structure note
Pages 1-6
DOI 10.1007/s10858-012-9608-7
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02-13-2012 02:34 AM
[NMR paper] Two-dimensional 1H NMR study of recombinant insect defensin A in water: resonance ass
Two-dimensional 1H NMR study of recombinant insect defensin A in water: resonance assignments, secondary structure and global folding.
Related Articles Two-dimensional 1H NMR study of recombinant insect defensin A in water: resonance assignments, secondary structure and global folding.
J Biomol NMR. 1992 May;2(3):235-56
Authors: Bonmatin JM, Bonnat JL, Gallet X, Vovelle F, Ptak M, Reichhart JM, Hoffmann JA, Keppi E, Legrain M, Achstetter T
A 500 MHz 2D 1H NMR study of recombinant insect defensin A is reported. This defense protein of 40...
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08-21-2010 11:41 PM
[NMR paper] Comparison of black coral skeleton and insect cuticle by a combination of carbon-13 N
Comparison of black coral skeleton and insect cuticle by a combination of carbon-13 NMR and chemical analyses.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Comparison of black coral skeleton and insect cuticle by a combination of carbon-13 NMR and chemical analyses.
Arch Biochem Biophys. 1992 Jan;292(1):107-11
Authors: Holl SM, Schaefer J, Goldberg WM, Kramer KJ, Morgan TD, Hopkins TL
Cross-polarization, magic-angle spinning 13C NMR spectra of skeletal components of...
Solution structure of insect CSP and OBPs by NMR.
Linear Mode
