title: Cloning, expression, crystallization and preliminary X-ray studies of a superfolder GFP fusion of cyanobacterial Psb32.
abstract: A fusion of Psb32 from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 (TePsb32) with superfolder GFP was created for enhanced solubility and improved detection and purification. The fusion protein readily formed large hexagonal crystals belonging to space group P6₁22. A full data set extending to 2.3 Å resolution was collected at the Swiss Light Source. The phase problem could be solved by using only the sfGFP fusion partner or by using GFP and AtTLP18.3 from Arabidopsis thaliana as search models. Based on this expression construct, a versatile library of 24 vectors combining four different superfolder GFP variants and three affinity tags was generated to facilitate expression and screening of fluorescent fusion proteins.
authors: Department of Plant Biochemistry, Ruhr-University Bochum, 44780 Bochum, Germany.
authors: Pasqual
authors: P
authors: Liauw
authors: Department of Plant Biochemistry, Ruhr-University Bochum, 44780 Bochum, Germany.
authors: Daniela
authors: D
authors: Kannchen
authors: Department of Biophysics, Ruhr-University Bochum, 44780 Bochum, Germany.
authors: Raphael
authors: R
authors: Gasper
authors: Department of Plant Biochemistry, Ruhr-University Bochum, 44780 Bochum, Germany.
authors: Nina
authors: N
authors: Dyczmons-Nowaczyk
authors: Department of Plant Biochemistry, Ruhr-University Bochum, 44780 Bochum, Germany.
authors: Marc M
authors: MM
authors: Nowaczyk
authors: Department of Biophysics, Ruhr-University Bochum, 44780 Bochum, Germany.
authors: Eckhard
authors: E
authors: Hofmann
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods; D003460:Crystallization; D018360:Crystallography, X-Ray; D000458:Cyanobacteria / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D050576:Membrane Fusion Proteins / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D017510:Protein Folding
chemical_list: D050576:Membrane Fusion Proteins
keywords: Psb32; Thermosynechococcus elongatus BP-1; superfolder GFP