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Cloning, expression, crystallization and crystallographic analysis of CouR from Rhodopseudomonas palustris
Author(s) -
Pan Chen,
Hu Yonglin,
Jiang Xiangning,
Gai Ying
Publication year - 2015
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/s2053230x15018968
Subject(s) - ligand (biochemistry) , crystallization , rhodopseudomonas palustris , stereochemistry , crystallography , dna , chemistry , cloning (programming) , biology , receptor , biochemistry , genetics , organic chemistry , bacteria , computer science , programming language
CouR from Rhodopseudomonas palustris is a member of the MarR transcriptional regulator family. It regulates the expression of CouA and CouB, enzymes that are involved in the degradation of p ‐coumarate. In vivo , CouR binds to a DNA fragment containing the couAB promoter and suppresses the expression of CouA and CouB, while binding of p ‐coumaroyl‐CoA attenuates its affinity towards DNA and activates the expression of CouA and CouB. Here, the crystallization and X‐ray diffraction analyses of CouR alone and in complex with p ‐coumaroyl‐CoA are reported. Apo and ligand‐complexed CouR crystals diffracted to 2.5 and 3.3 Å resolution, respectively. The crystals of apo CouR belonged to space group P 22 1 2 1 , with unit‐cell parameters a = 62.78, b = 76.15, c = 87.38 Å, whereas the crystals of the CouR–ligand complex belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 61.37, b = 69.82, c = 70.32 Å. The crystals were predicted to contain two CouR molecules or CouR–ligand complexes per asymmetric unit.