Protein expression, characterization, crystallization and preliminary X-ray crystallographic analysis of a Fic protein from Clostridium difficile

Ditte Welner, Emil Dedic, Hans C. Van Leeuwen, Ed Kuijper, Morten Jannik Bjerrum, Ole Østergaard, René Jørgensen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Fic domains in proteins are found in abundance in nature from the simplest prokaryotes to animals. Interestingly, Fic domains found in two virulence factors of Gram-negative bacteria have recently been demonstrated to catalyse the transfer of the AMP moiety from ATP to small host GTPases. This post-translational modification has attracted considerable interest and a role for adenylylation in pathology and physiology is emerging. This work was aimed at the structural characterization of a newly identified Fic protein of the Gram-positive bacterium Clostridium difficile. A constitutively active inhibitory helix mutant of C. difficile Fic was overexpressed in Escherichia coli, purified and crystallized by the vapour-diffusion technique. Preliminary X-ray crystallographic analysis shows that the crystals diffract to at least 1.68 Å resolution at a synchrotron X-ray source. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 45.6, b = 80.8, c = 144.7 Å, α = β = γ = 90°. Two molecules per asymmetric unit corresponds to a Matthews coefficient of 2.37 Å3 Da-1 and a solvent content of 48%.

Original languageEnglish
JournalActa Crystallographica Section F:Structural Biology Communications
Volume70
Issue number6
Pages (from-to)827-831
Number of pages5
DOIs
Publication statusPublished - Jun 2014
Externally publishedYes

Keywords

  • adenylylation
  • Clostridium difficile
  • Fic protein

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