Purification, crystallization and preliminary X-ray diffraction analysis of the 23S rRNA methyltransferase RlmJ from Escherichia coli

Avinash S. Punekar, Maria Selmer (Lead / Corresponding author)

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Methyltransferase RlmJ uses the cofactor S-adenosylmethionine to methylate the exocyclic nitrogen N6 of nucleotide A2030 in 23S rRNA during ribosome assembly in Escherichia coli. RlmJ with a C-terminal hexahistidine tag was overexpressed in E. coli 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 P2(1), with unit-cell parameters a = 46.9, b = 77.8, c = 82.5 Å, β = 104°. Data analysis suggested two molecules per asymmetric unit and a Matthews coefficient of 2.20 Å3 Da-1.

Original languageEnglish
Pages (from-to)1001-1003
Number of pages3
JournalActa Crystallographica F-Structural Biology and Crystallization Communications
Volume69
Issue numberPt 9
DOIs
Publication statusPublished - Sep 2013

Keywords

  • Amino Acid Sequence
  • Crystallography, X-Ray
  • Escherichia coli/chemistry
  • Escherichia coli Proteins/chemistry
  • Gene Expression
  • Methyltransferases/chemistry
  • Molecular Sequence Data
  • Protein Interaction Domains and Motifs
  • RNA, Ribosomal, 23S/chemistry
  • Recombinant Fusion Proteins/chemistry
  • Ribosomes/genetics
  • S-Adenosylmethionine/chemistry

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