title: Expression, purification, crystallization and preliminary X-ray analysis of eCGP123, an extremely stable monomeric green fluorescent protein with reversible photoswitching properties.
abstract: 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 P1, 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Biomedical Science, Monash University, Clayton Campus, Melbourne, Victoria 3800, Australia.
authors.forename: Craig
authors.initials: C
authors.lastname: Don Paul
authors.forename: Daouda A K
authors.initials: DA
authors.lastname: Traore
authors.forename: Emma
authors.initials: E
authors.lastname: Byres
authors.forename: Jamie
authors.initials: J
authors.lastname: Rossjohn
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Devenish
authors.forename: Csaba
authors.initials: C
authors.lastname: Kiss
authors.forename: Andrew
authors.initials: A
authors.lastname: Bradbury
authors.forename: Matthew C J
authors.initials: MC
authors.lastname: Wilce
authors.forename: Mark
authors.initials: M
authors.lastname: Prescott
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D055668:Photochemical Processes; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins
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