Open AccessCrystallization of recombinant bifunctional nuclease TBN1 from tomato
Author(s) -
Koval' Tomáš,
Lipovová Petra,
Podzimek Tomáš,
Matoušek Jaroslav,
Dušková Jarmila,
Skálová Tereza,
Štěpánková Andrea,
Hašek Jindřich,
Dohnálek Jan
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110048177
Subject(s) - orthorhombic crystal system , crystallography , nuclease , crystallization , solanum , crystal (programming language) , nicotiana benthamiana , resolution (logic) , protoplast , chemistry , materials science , crystal structure , biology , botany , biochemistry , enzyme , gene , organic chemistry , artificial intelligence , computer science , programming language
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 Zn 2+ 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 Zn 2+ ions in a similar spatial arrangement to that observed in nuclease P1 from Penicillium citrinum .