title: Expression, purification, crystallization and preliminary X-ray crystallographic analysis of a novel plant-type ferredoxin/thioredoxin reductase-like protein from Methanosarcina acetivorans.
abstract: 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 P23 or P2(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 (VM) of 3.55 Å3 Da(-1), corresponding to a solvent content of 65%.
authors.affiliation: Biochemistry and Molecular Biology, Pennsylvania State University, PA 16802, USA.
authors.forename: Adepu K
authors.initials: AK
authors.lastname: Kumar
authors.forename: Neela H
authors.initials: NH
authors.lastname: Yennawar
authors.forename: Hemant P
authors.initials: HP
authors.lastname: Yennawar
authors.forename: James G
authors.initials: JG
authors.lastname: Ferry
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D017020:Methanosarcina / Q000737:chemistry*; D008969:Molecular Sequence Data; D012416:Rubredoxins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment
chemical_list: D019843:Archaeal Proteins; D012416:Rubredoxins