title: Crystallization of recombinant bifunctional nuclease TBN1 from tomato.
abstract: The endonuclease TBN1 from Solanum lycopersicum (tomato) was expressed in Nicotiana benthamiana leaves and purified with suitable quality and in suitable quantities for crystallization experiments. Two crystal forms (orthorhombic and rhombohedral) were obtained and X-ray diffraction experiments were performed. The presence of natively bound Zn2+ ions was confirmed by X-ray fluorescence and by an absorption-edge scan. X-ray diffraction data were collected from the orthorhombic (resolution of 5.2 Å) and rhombohedral (best resolution of 3.2 Å) crystal forms. SAD, MAD and MR methods were applied for solution of the phase problem, with partial success. TBN1 contains three Zn2+ ions in a similar spatial arrangement to that observed in nuclease P1 from Penicillium citrinum.
authors.affiliation: Institute of Physics AS CR, v.v.i., Na Slovance 2, 182 21 Praha 8, Czech Republic. koval.tomas@gmail.com
authors.forename: Tomáš
authors.initials: T
authors.lastname: Koval'
authors.forename: Petra
authors.initials: P
authors.lastname: Lipovová
authors.forename: Tomáš
authors.initials: T
authors.lastname: Podzimek
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Matoušek
authors.forename: Jarmila
authors.initials: J
authors.lastname: Dušková
authors.forename: Tereza
authors.initials: T
authors.lastname: Skálová
authors.forename: Andrea
authors.initials: A
authors.lastname: Stěpánková
authors.forename: Jindřich
authors.initials: J
authors.lastname: Hašek
authors.forename: Jan
authors.initials: J
authors.lastname: Dohnálek
mesh_terms: D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000235:genetics; D007477:Ions / Q000737:chemistry; D018551:Solanum lycopersicum / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics; D015032:Zinc / Q000737:chemistry
chemical_list: D007477:Ions; D010940:Plant Proteins; D011994:Recombinant Proteins; D003851:Deoxyribonucleases; D015032:Zinc