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
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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. / Stella, Stefano; Molina, Rafael; Bertonatti, Claudia; Juillerrat, Alexandre; Montoya, Guillermo.
In: Acta Crystallographica Section F:Structural Biology Communications, Vol. 70, No. 1, 01.01.2014, p. 87-91.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - 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
AU - Stella, Stefano
AU - Molina, Rafael
AU - Bertonatti, Claudia
AU - Juillerrat, Alexandre
AU - Montoya, Guillermo
PY - 2014/1/1
Y1 - 2014/1/1
N2 - 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 P31 or its enantiomorph P32, 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 P21, 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.
AB - 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 P31 or its enantiomorph P32, 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 P21, 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.
KW - Burkholderia rhizoxinica
KW - BurrH
KW - gene targeting
KW - protein-DNA interaction
UR - http://www.scopus.com/inward/record.url?scp=84901403172&partnerID=8YFLogxK
U2 - 10.1107/S2053230X13033037
DO - 10.1107/S2053230X13033037
M3 - Journal article
C2 - 24419625
AN - SCOPUS:84901403172
VL - 70
SP - 87
EP - 91
JO - Acta Crystallographica Section F: Structural Biology Communications
JF - Acta Crystallographica Section F: Structural Biology Communications
SN - 2053-230X
IS - 1
ER -
ID: 202332631