Open AccessExpression, purification, crystallization and preliminary X‐ray crystallographic analysis of a novel plant‐type ferredoxin/thioredoxin reductase‐like protein from Methanosarcina acetivorans
Author(s) -
Kumar Adepu K.,
Yennawar Neela H.,
Yennawar Hemant P.,
Ferry James G.
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111017234
Subject(s) - ferredoxin , crystallization , ferredoxin thioredoxin reductase , crystallography , reductase , methanosarcina barkeri , chemistry , thioredoxin , thioredoxin reductase , biochemistry , biology , enzyme , genetics , organic chemistry , methanogenesis , bacteria
The genome of Methanosarcina acetivorans contains a gene ( ma1659 ) that is predicted to encode an uncharacterized chimeric protein containing a plant‐type ferredoxin/thioredoxin reductase‐like catalytic domain in the N‐terminal region and a bacterial‐like rubredoxin domain in the C‐terminal region. To understand the structural and functional properties of the protein, the ma1659 gene was cloned and overexpressed in Escherichia coli . Crystals of the MA1659 protein were grown by the sitting‐drop method using 2 M ammonium sulfate, 0.1 M HEPES buffer pH 7.5 and 0.1 M urea. Diffraction data were collected to 2.8 Å resolution using the remote data‐collection feature of the Advanced Light Source, Lawrence Berkeley National Laboratory. The crystal belonged to the primitive cubic space group P 23 or P 2 1 3, with unit‐cell parameters a = b = c = 92.72 Å. Assuming the presence of one molecule in the asymmetric unit gave a Matthews coefficient ( V M ) of 3.55 Å 3 Da −1 , corresponding to a solvent content of 65%.