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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.

SUBMITTER: Koval' T

PROVIDER: S-EPMC3079990 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature

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Crystallization of recombinant bifunctional nuclease TBN1 from tomato.

Koval' Tomáš T Lipovová Petra P Podzimek Tomáš T Matoušek Jaroslav J Dušková Jarmila J Skálová Tereza T Stěpánková Andrea A Hašek Jindřich J Dohnálek Jan J

Acta crystallographica. Section F, Structural biology and crystallization communications 20101223 Pt 1

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 ...[more]

PMID: 21206042

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