Parallel screening and optimization of protein constructs for structural studies.
A major challenge in structural biology remains the identification of protein constructs amenable to structural characterization. Here, we present a simple method for parallel expression, labeling, and purification of protein constructs (up to 80 kDa) combined with rapid evaluation by NMR spectroscopy. Our approach, which is equally applicable for manual or automated implementation, offers an efficient way to identify and optimize protein constructs for NMR or X-ray crystallographic investigations.
Rasia
Rodolfo M
RM
Institut de Biologie Structurale Jean-Pierre Ebel, Centre National de la Recherche Scientifique, Université Joseph-Fourier, Grenoble, France.
Noirclerc-Savoye
Marjolaine
M
Bologna
Nicolás G
NG
Gallet
Benoit
B
Plevin
Michael J
MJ
Blanchard
Laurence
L
Palatnik
Javier F
JF
Brutscher
Bernhard
B
Vernet
Thierry
T
Boisbouvier
Jérôme
J
D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D007553:Isotope Labeling; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D029681:Arabidopsis Proteins; C420966:HYL1 protein, Arabidopsis; D009587:Nitrogen Isotopes; D011506:Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins