Refolding, characterization, and preliminary X-ray crystallographic studies on the campylobacter concisus plasmid-encoded secreted protein Csep1p associated with Crohn’s disease

Mohammad Mizanur Rahman, Bradley Goff, Li Zhang, Anna Roujeinikova

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Colonization of Campylobacter concisus in the gastrointestinal tract can lead to the development of inflammatory bowel disease (IBD). Plasmid-encoded C. concisus-secreted protein 1 (Csep1p) was recently identified as a putative pathogenicity marker associated with active Crohn’s disease, a clinical form of IBD. Csep1p shows no significant full-length sequence similarity to proteins of known structure, and its role in pathogenesis is not yet known. This study reports a method for extraction of recombinantly expressed Csep1p from Escherichia coliinclusion bodies, refolding, and purification to produce crystallizable protein. Purified recombinant Csep1p behaved as a monomer in solution. Crystals of Csep1p were grown by the hanging drop vapour diffusion method, using polyethylene glycol (PEG) 4000 as the precipitating agent. A complete data set has been collected to 1.4 Å resolution, using cryocooling conditions and synchrotron radiation. The crystals belong to space group P62 or P64, with unit cell parameters a = b = 85.8, c = 55.2 Å, α = β = 90, and γ = 120°. The asymmetric unit appears to contain one subunit, corresponding to a packing density of 2.47 Å3 Da−1.

Original languageEnglish
Article number391
Number of pages10
JournalCrystals
Volume8
Issue number10
DOIs
Publication statusPublished - 16 Oct 2018

Keywords

  • Campylobacter consisus
  • Circular dichroism
  • Crohn’s disease
  • Csep1
  • Protein crystallization

Cite this

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title = "Refolding, characterization, and preliminary X-ray crystallographic studies on the campylobacter concisus plasmid-encoded secreted protein Csep1p associated with Crohn’s disease",
abstract = "Colonization of Campylobacter concisus in the gastrointestinal tract can lead to the development of inflammatory bowel disease (IBD). Plasmid-encoded C. concisus-secreted protein 1 (Csep1p) was recently identified as a putative pathogenicity marker associated with active Crohn’s disease, a clinical form of IBD. Csep1p shows no significant full-length sequence similarity to proteins of known structure, and its role in pathogenesis is not yet known. This study reports a method for extraction of recombinantly expressed Csep1p from Escherichia coliinclusion bodies, refolding, and purification to produce crystallizable protein. Purified recombinant Csep1p behaved as a monomer in solution. Crystals of Csep1p were grown by the hanging drop vapour diffusion method, using polyethylene glycol (PEG) 4000 as the precipitating agent. A complete data set has been collected to 1.4 {\AA} resolution, using cryocooling conditions and synchrotron radiation. The crystals belong to space group P62 or P64, with unit cell parameters a = b = 85.8, c = 55.2 {\AA}, α = β = 90, and γ = 120°. The asymmetric unit appears to contain one subunit, corresponding to a packing density of 2.47 {\AA}3 Da−1.",
keywords = "Campylobacter consisus, Circular dichroism, Crohn’s disease, Csep1, Protein crystallization",
author = "Rahman, {Mohammad Mizanur} and Bradley Goff and Li Zhang and Anna Roujeinikova",
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Refolding, characterization, and preliminary X-ray crystallographic studies on the campylobacter concisus plasmid-encoded secreted protein Csep1p associated with Crohn’s disease. / Rahman, Mohammad Mizanur; Goff, Bradley; Zhang, Li; Roujeinikova, Anna.

In: Crystals, Vol. 8, No. 10, 391, 16.10.2018.

Research output: Contribution to journalArticleResearchpeer-review

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