Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α-carbonic anhydrase with acetazolamide

Joyanta K. Modak, Sarah A. Revitt-Mills, Anna Roujeinikova

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Helicobacter pylori infection of the stomach can lead to severe gastroduodenal diseases such as gastritis, peptic ulcers and gastric cancers. Periplasmic H. pylori α-carbonic anhydrase (HpαCA) is essential for the acclimatization of the bacterium to the acidity of the stomach. Through the action of urease and carbonic anhydrases, the H. pylori periplasmic pH is maintained at around 6 in an environment with a pH as low as 2, which in turn facilitates the maintenance of a cytoplasmic pH close to neutral, allowing growth in the gastric niche. Crystals of HpαCA in complex with the inhibitor acetazolamide have been grown by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitating agent. The crystals have the symmetry of space group P212121, with unit-cell parameters a = 37.0, b = 82.4, c = 150.8 Å. An X-ray diffraction data set was collected from a single crystal to 1.7 Å resolution. Calculation of the self-rotation function using this data and molecular replacement showed that the asymmetric unit contains an HpαCA dimer.
Original languageEnglish
Pages (from-to)1252-1255
Number of pages4
JournalActa Crystallographica. Section F: Structural Biology Communications
VolumeF69
Issue number11
DOIs
Publication statusPublished - 2013

Keywords

  • helicobacter pylori
  • alpha-carbonic anhydrase
  • acetazolamide

Cite this

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title = "Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α-carbonic anhydrase with acetazolamide",
abstract = "Helicobacter pylori infection of the stomach can lead to severe gastroduodenal diseases such as gastritis, peptic ulcers and gastric cancers. Periplasmic H. pylori α-carbonic anhydrase (HpαCA) is essential for the acclimatization of the bacterium to the acidity of the stomach. Through the action of urease and carbonic anhydrases, the H. pylori periplasmic pH is maintained at around 6 in an environment with a pH as low as 2, which in turn facilitates the maintenance of a cytoplasmic pH close to neutral, allowing growth in the gastric niche. Crystals of HpαCA in complex with the inhibitor acetazolamide have been grown by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitating agent. The crystals have the symmetry of space group P212121, with unit-cell parameters a = 37.0, b = 82.4, c = 150.8 {\AA}. An X-ray diffraction data set was collected from a single crystal to 1.7 {\AA} resolution. Calculation of the self-rotation function using this data and molecular replacement showed that the asymmetric unit contains an HpαCA dimer.",
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Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α-carbonic anhydrase with acetazolamide. / Modak, Joyanta K.; Revitt-Mills, Sarah A.; Roujeinikova, Anna.

In: Acta Crystallographica. Section F: Structural Biology Communications, Vol. F69, No. 11, 2013, p. 1252-1255.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α-carbonic anhydrase with acetazolamide

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