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Expression, purification, crystallization and preliminary X‐ray diffraction analysis of the novel modular DNA‐binding protein BurrH in its apo form and in complex with its target DNA
Author(s) -
Stella Stefano,
Molina Rafael,
Bertonatti Claudia,
Juillerrat Alexandre,
Montoya Guillermo
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x13033037
Subject(s) - monoclinic crystal system , crystallography , crystallization , dna , resolution (logic) , chemistry , diffraction , x ray crystallography , molecule , crystal structure , biochemistry , physics , organic chemistry , artificial intelligence , computer science , optics
Different genome‐editing strategies have fuelled the development of new DNA‐targeting molecular tools allowing precise gene modifications. Here, the expression, purification, crystallization and preliminary X‐ray diffraction of BurrH, a novel DNA‐binding protein from Burkholderia rhizoxinica , are reported. Crystallization experiments of BurrH in its apo form and in complex with its target DNA yielded crystals suitable for X‐ray diffraction analysis. The crystals of the apo form belonged to the primitive hexagonal space group P 3 1 or its enantiomorph P 3 2 , with unit‐cell parameters a = b = 73.28, c = 268.02 Å, α = β = 90, γ = 120°. The BurrH–DNA complex crystallized in the monoclinic space group P 2 1 , with unit‐cell parameters a = 70.15, b = 95.83, c = 76.41 Å, α = γ = 90, β = 109.51°. The self‐rotation function and the Matthews coefficient suggested the presence of two protein molecules per asymmetric unit in the apo crystals and one protein–DNA complex in the monoclinic crystals. The crystals diffracted to resolution limits of 2.21 and 2.65 Å, respectively, using synchrotron radiation.