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Crystallization and preliminary X‐ray crystallographic analysis of carboxyl‐terminal region 4 of SigR from Streptomyces coelicolor A3(2)
Author(s) -
Kim Keon Young,
Kim Sunmin,
Park Jeong Kuk,
Song HyoJin,
Park SangYoun
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/s2053230x14008437
Subject(s) - streptomyces coelicolor , crystallization , crystallography , crystal (programming language) , polyethylene glycol , x ray , resolution (logic) , crystal structure , chemistry , protein crystallization , x ray crystallography , powder diffraction , diffraction , materials science , physics , optics , organic chemistry , biochemistry , artificial intelligence , computer science , mutant , gene , programming language
Full‐length SigR from Streptomyces coelicolor A3(2) was overexpressed in Escherichia coli , purified and submitted to crystallization trials using either polyethylene glycol 3350 or 4000 as a precipitant. X‐ray diffraction data were collected to 2.60 Å resolution under cryoconditions using synchrotron X‐rays. The crystal packs in space group P 4 3 2 1 2, with unit‐cell parameters a = b = 42.14, c = 102.02 Å. According to the Matthews coefficient, the crystal asymmetric unit cannot contain the full‐length protein. Molecular replacement with the known structures of region 2 and region 4 as independent search models indicates that the crystal contains only the −35 element‐binding carboxyl‐terminal region 4 of full‐length SigR. Mass‐spectrometric analysis of the harvested crystal confirms this, suggesting a crystal volume per protein weight ( V M ) of 2.24 Å 3 Da −1 and 45.1% solvent content.