Expression, purification and preliminary crystallographic analysis of the Toxoplasma gondii enoyl reductase

Stephen P. Muench, Sean Taylor Prigge, Liqun Zhu, Michael J. Kirisits, Craig W. Roberts, Sarah Wernimont, Rima McLeod, David W. Rice

Research output: Contribution to journalArticle

Abstract

The protozoan parasite Toxoplasma gondii is the causative agent of one of the most widespread parasitic infections of man and is a leading cause of congenital neurological birth defects and the third most common cause of food-borne deaths in the United States. Despite this, to date no drugs are available that provide a fully effective treatment. Recently, the antibacterial agent triclosan was shown to inhibit the fatty-acid biosynthesis pathway in T. gondii and to interact with the enoyl reductase (ENR). In order to analyse the potential of triclosan as a lead compound targeting T. gondii ENR and to explore unique features of the apicomplexan enzyme that could be exploited in future drug development, structural studies have been initiated on T. gondii ENR. Crystals of T. gondii ENR in complex with NAD+ and triclosan were grown using the hanging-drop vapour-diffusion method with PEG 8000 as precipitant. The crystals belong to space group P3221, with approximate unit-cell parameters a = 78.1, b = 78.1, c = 188.5 Å, α = β = 90, γ = 120° and a dimer in the asymmetric unit. Test data were collected to beyond 2.6 Å on cryocooled crystals (100 K) using a Rigaku MM007 rotating-anode X-ray source, revealing that the crystals are suitable for a full structural determination.

Original languageEnglish (US)
Pages (from-to)604-606
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume62
Issue number6
DOIs
StatePublished - Jun 2006

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Toxoplasma
Triclosan
purification
Purification
Oxidoreductases
Crystals
crystals
drugs
Lead compounds
lead compounds
parasites
biosynthesis
causes
Biosynthesis
infectious diseases
fatty acids
Parasitic Diseases
food
death
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Structural Biology
  • Genetics
  • Condensed Matter Physics

Cite this

Expression, purification and preliminary crystallographic analysis of the Toxoplasma gondii enoyl reductase. / Muench, Stephen P.; Prigge, Sean Taylor; Zhu, Liqun; Kirisits, Michael J.; Roberts, Craig W.; Wernimont, Sarah; McLeod, Rima; Rice, David W.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 62, No. 6, 06.2006, p. 604-606.

Research output: Contribution to journalArticle

Muench, Stephen P. ; Prigge, Sean Taylor ; Zhu, Liqun ; Kirisits, Michael J. ; Roberts, Craig W. ; Wernimont, Sarah ; McLeod, Rima ; Rice, David W. / Expression, purification and preliminary crystallographic analysis of the Toxoplasma gondii enoyl reductase. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2006 ; Vol. 62, No. 6. pp. 604-606.
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