Open AccessExpression, purification, crystallization and preliminary X‐ray analysis of eCGP123, an extremely stable monomeric green fluorescent protein with reversible photoswitching properties
Author(s) -
Don Paul Craig,
Traore Daouda A. K.,
Byres Emma,
Rossjohn Jamie,
Devenish Rodney J.,
Kiss Csaba,
Bradbury Andrew,
Wilce Matthew C. J.,
Prescott Mark
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111028156
Subject(s) - green fluorescent protein , crystallization , fluorescence , peg ratio , monomer , protein crystallization , diffraction , solvent , crystallography , molecule , resolution (logic) , x ray crystallography , chemistry , crystal (programming language) , x ray , materials science , polymer , optics , organic chemistry , biochemistry , physics , finance , artificial intelligence , computer science , economics , gene , programming language
Enhanced consensus green protein variant 123 (eCGP123) is an extremely thermostable green fluorescent protein (GFP) that exhibits useful negative reversible photoswitching properties. eCGP123 was derived by the application of both a consensus engineering approach and a recursive evolutionary process. Diffraction‐quality crystals of recombinant eCGP123 were obtained by the hanging‐drop vapour‐diffusion method using PEG 3350 as the precipitant. The eCGP123 crystal diffracted X‐rays to 2.10 Å resolution. The data were indexed in space group P 1, with unit‐cell parameters a = 74.63, b = 75.38, c = 84.51 Å, α = 90.96, β = 89.92, γ = 104.03°. The Matthews coefficient ( V M = 2.26 Å 3 Da −1 ) and a solvent content of 46% indicated that the asymmetric unit contained eight eCGP123 molecules.