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Cloning, expression, purification, crystallization and preliminary X-ray diffraction data of the Pyrococcus horikoshii RadA intein.

ABSTRACT: The RadA intein from the hyperthermophilic archaebacterium Pyrococcus horikoshii was cloned, expressed and purified for subsequent structure determination. The protein crystallized rapidly in several conditions. The best crystals, which diffracted to 1.75 Å resolution, were harvested from drops consisting of 0.1 M HEPES pH 7.5, 3.0 M NaCl and were cryoprotected with Paratone-N before flash-cooling. The collected data were processed in the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 58.1, b = 67.4, c = 82.9 Å. Molecular replacement with Rosetta using energy- and density-guided structure optimization provided the initial solution, which is currently under refinement.

SUBMITTER: Lyskowski A

PROVIDER: S-EPMC3087655 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature

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Cloning, expression, purification, crystallization and preliminary X-ray diffraction data of the Pyrococcus horikoshii RadA intein.

Lyskowski Andrzej A Oeemig Jesper S JS Jaakkonen Anniina A Rommi Katariina K DiMaio Frank F Zhou Dongwen D Kajander Tommi T Baker David D Wlodawer Alexander A Goldman Adrian A Iwaï Hideo H

Acta crystallographica. Section F, Structural biology and crystallization communications 20110428 Pt 5

The RadA intein from the hyperthermophilic archaebacterium Pyrococcus horikoshii was cloned, expressed and purified for subsequent structure determination. The protein crystallized rapidly in several conditions. The best crystals, which diffracted to 1.75 Å resolution, were harvested from drops consisting of 0.1 M HEPES pH 7.5, 3.0 M NaCl and were cryoprotected with Paratone-N before flash-cooling. The collected data were processed in the orthorhombic space group P2(1)2(1)2(1), with unit-cell pa ...[more]

PMID: 21543876

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