Open AccessCloning, expression, purification, crystallization and preliminary X‐ray analysis of the human RuvBL1–RuvBL2 complex
Author(s) -
Gorynia Sabine,
Matias Pedro M.,
Bandeiras Tiago M.,
Donner Peter,
Carrondo Maria Arménia
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910802558x
Subject(s) - cloning (programming) , crystallization , materials science , microbiology and biotechnology , virology , chemistry , biology , computer science , programming language , organic chemistry
The complex of RuvBL1 and its homologue RuvBL2, two evolutionarily highly conserved eukaryotic proteins belonging to the AAA + (ATPase associated with diverse cellular activities) family of ATPases, was co‐expressed in Escherichia coli . For crystallization purposes, the flexible domains II of RuvBL1 and RuvBL2 were truncated. The truncated RuvBL1–RuvBL2 complex was crystallized using the hanging‐drop vapour‐diffusion method at 293 K. The crystals were hexagonal‐shaped plates and belonged to either the orthorhombic space group C 222 1 , with unit‐cell parameters a = 111.4, b = 188.0, c = 243.4 Å and six monomers in the asymmetric unit, or the monoclinic space group P 2 1 , with unit‐cell parameters a = 109.2, b = 243.4, c = 109.3 Å, β = 118.7° and 12 monomers in the asymmetric unit. The crystal structure could be solved by molecular replacement in both possible space groups and the solutions obtained showed that the complex forms a dodecamer.