In-cell NMR provides structural and functional information on proteins directly inside living cells. At present, the high costs of the labeled media for mammalian cells represent a limiting factor for the development of this methodology. Here we report a protocol to prepare a homemade growth medium from Spirulina platensis autolysate, suitable to express uniformly labeled proteins inside mammalian cells at a reduced cost-per-sample. The human proteins SOD1 and Mia40 were overexpressed in human cells grown in 15N-enriched S. platensis algal-derived medium, and high quality in-cell NMR spectra were obtained.
[NMR paper] Cell-free expression of the APP transmembrane fragments with Alzheimer's disease mutations using algal amino acid mixture for structural NMR studies.
Cell-free expression of the APP transmembrane fragments with Alzheimer's disease mutations using algal amino acid mixture for structural NMR studies.
Related Articles Cell-free expression of the APP transmembrane fragments with Alzheimer's disease mutations using algal amino acid mixture for structural NMR studies.
Protein Expr Purif. 2016 Apr 9;
Authors: Bocharova OV, Urban AS, Nadezhdin KD, Bocharov EV, Arseniev AS
Abstract
Structural investigations need ready supply of the isotope labeled proteins with inserted mutations n the...
nmrlearner
Journal club
0
04-14-2016 12:01 PM
[NMR paper] Fluorine NMR-based assay in living mammalian cells.
Fluorine NMR-based assay in living mammalian cells.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Fluorine NMR-based assay in living mammalian cells.
Anal Biochem. 2015 Dec 11;
Authors: Veronesi M, Giacomina F, Romeo E, Castellani B, Ottonello G, Lambruschini C, Garau G, Scarpelli R, Bandiera T, Piomelli D, Dalvit C
Abstract
Nuclear Magnetic Resonance (NMR)-based screening has been recognized as a powerful approach for the identification...
nmrlearner
Journal club
0
12-28-2015 12:26 AM
A PositivelyCharged Liquid Crystalline Medium forMeasuring Residual Dipolar Couplings in Membrane Proteins by NMR
A PositivelyCharged Liquid Crystalline Medium forMeasuring Residual Dipolar Couplings in Membrane Proteins by NMR
Pallavi Thiagarajan-Rosenkranz, Adrian W. Draney, Sean T. Smrt and Justin L. Lorieau
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/jacs.5b07515/20150910/images/medium/ja-2015-07515m_0004.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.5b07515
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/Q6RR1Z4u0gw
nmrlearner
Journal club
0
09-11-2015 06:48 AM
[NMR paper] A Positively Charged Liquid Crystalline Medium for Measuring Residual Dipolar Couplings in Membrane Proteins by NMR.
A Positively Charged Liquid Crystalline Medium for Measuring Residual Dipolar Couplings in Membrane Proteins by NMR.
A Positively Charged Liquid Crystalline Medium for Measuring Residual Dipolar Couplings in Membrane Proteins by NMR.
J Am Chem Soc. 2015 Sep 8;
Authors: Thiagarajan-Rosenkranz Paltr Uic Edu P, Draney AW, Smrt ST, Lorieau JL
Abstract
Residual Dipolar Couplings (RDCs) are integral to the refinement of membrane protein structures by NMR since they accurately define the orientation of helices and other structural...
nmrlearner
Journal club
0
09-09-2015 11:49 AM
[NMR paper] Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
J Struct Biol. 2014 Aug 27;
Authors: Meola A, Deville C, Jeffers SA, Guardado-Calvo P, Vasiliauskaite I, Sizun C, Girard-Blanc C, Malosse C, Heijenoort CV, Chamot-Rooke J, Krey T, Guittet E, Pêtres S, Rey FA, Bontems F
Abstract
Nuclear...
nmrlearner
Journal club
0
09-01-2014 07:46 PM
Robust and low cost uniform 15N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications
Robust and low cost uniform 15N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications
Publication date: Available online 28 August 2014
Source:Journal of Structural Biology</br>
Author(s): Annalisa Meola , Célia Deville , Scott A. Jeffers , Pablo Guardado-Calvo , Ieva Vasiliauskaite , Christina Sizun , Christine Girard-Blanc , Christian Malosse , Carine van Heijenoort , Julia Chamot-Rooke , Thomas Krey , Eric Guittet , Stéphane Pêtres , Félix A. Rey , François Bontems</br>
Nuclear magnetic...
nmrlearner
Journal club
0
08-29-2014 05:36 PM
[NMR paper] Using MUSIC and CC(CO)NH for Backbone Assignment of Two Medium-Sized Proteins Not Fully Accessible to Standard 3D NMR.
Using MUSIC and CC(CO)NH for Backbone Assignment of Two Medium-Sized Proteins Not Fully Accessible to Standard 3D NMR.
Related Articles Using MUSIC and CC(CO)NH for Backbone Assignment of Two Medium-Sized Proteins Not Fully Accessible to Standard 3D NMR.
Molecules. 2014;19(7):8890-8903
Authors: Brenner AK, Frøystein NA
Abstract
The backbone assignment of medium-sized proteins is rarely as straightforward as that of small proteins, and thus often requires creative solutions. Here, we describe the application of a combination of...
nmrlearner
Journal club
0
06-29-2014 02:00 AM
[Question from NMRWiki Q&A forum] When to label peptides and proteins and when not?
When to label peptides and proteins and when not?
When pursuing an NMR structure project - at what size of oligopeptide you would decide to label it with:
15N
13C
both
Algal autolysate medium to label proteins for NMR in mammalian cells
Linear Mode
