Purification, crystallization and preliminary X‐ray diffraction analysis of the 23S rRNA methyltransferase RlmJ from Escherichia coli. Issue 9 (30th August 2013)

• Record Type: Journal Article
• Authors: Punekar, Avinash S.; Selmer, Maria
• Is Part Of: Acta crystallographica. Volume 69:Issue 9(2013:Sep.)
• Page Start: 1001
• Page End: 1003
• Publication Date: 2013-08-30
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309113020289
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Methyltransferase RlmJ uses the cofactor <italic>S</italic>‐adenosylmethionine to methylate the exocyclic nitrogen N6 of nucleotide A2030 in 23S rRNA during ribosome assembly in <italic>Escherichia coli</italic>. RlmJ with a C‐terminal hexahistidine tag was overexpressed in <italic>E. coli</italic> and purified as a monomer using Ni²⁺‐affinity and size‐exclusion chromatography. The recombinant RlmJ was crystallized using the sitting‐drop vapour‐diffusion method and a full data set was collected to 1.85 Å resolution from a single apo crystal. The crystals belonged to space group <italic>P</italic>2₁, with unit‐cell parameters <italic>a</italic> = 46.9, <italic>b</italic> = 77.8, <italic>c</italic> = 82.5 Å, β = 104°. Data analysis suggested two molecules per asymmetric unit and a Matthews coefficient of 2.20 ų Da⁻¹.</p> </abstract>