title: 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.
authors.affiliation: Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, PO Box 65, Helsinki FIN-00014, Finland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Lyskowski
authors.forename: Jesper S
authors.initials: JS
authors.lastname: Oeemig
authors.forename: Anniina
authors.initials: A
authors.lastname: Jaakkonen
authors.forename: Katariina
authors.initials: K
authors.lastname: Rommi
authors.forename: Frank
authors.initials: F
authors.lastname: DiMaio
authors.forename: Dongwen
authors.initials: D
authors.lastname: Zhou
authors.forename: Tommi
authors.initials: T
authors.lastname: Kajander
authors.forename: David
authors.initials: D
authors.lastname: Baker
authors.forename: Alexander
authors.initials: A
authors.lastname: Wlodawer
authors.forename: Adrian
authors.initials: A
authors.lastname: Goldman
authors.forename: Hideo
authors.initials: H
authors.lastname: Iwaï
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D008969:Molecular Sequence Data; D043902:Pyrococcus horikoshii / Q000737:chemistry*
chemical_list: D019843:Archaeal Proteins; D004268:DNA-Binding Proteins; C117252:RadA protein, archaeal